LOCUS Exported 6345 bp ds-DNA circular SYN 28-JUNI-2017 DEFINITION . ACCESSION . VERSION . KEYWORDS pOpt2_YFP_Ble SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6345) AUTHORS Thomas Baier TITLE Direct Submission JOURNAL Exported 6. Nov. 2017 from SnapGene 2.3.2 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6345 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(24..330) /direction=LEFT /label="f1ori" /note="f1ori" misc_feature complement(360..507) /label="lacZ_a_KS" /note="lacZ_a_KS" primer_bind complement(466..485) /label="Sequencing_primer_binding_site" /note="Sequencing_primer_binding_site" primer_bind complement(474..496) /label="M13_pUC-fwd_primer" /note="M13_pUC-fwd_primer" primer_bind complement(489..505) /label="M13_forward20_primer" /note="M13_forward20_primer" promoter 516..533 /label="T7_promoter" /note="T7_promoter" primer_bind 523..544 /label="InsrtCKpJR38_Fw" /note="InsrtCKpJR38_Fw" primer_bind 560..571 /label="pBluescriptSK_primer" /note="pBluescriptSK_primer" promoter 572..838 /label="HSP70Ap" /note="HSP70Ap promoter" promoter 845..1065 /label="RBCS2p" /note="RBCS2p promoter" transit_peptide join(1066..1073,1219) /note="RBCS2ORF" intron 1074..1218 /label="RBCS2i(1)" /note="RBCS2i" CDS 1247..1258 /codon_start=1 /product="Factor Xa recognition and cleavage site" /note="Factor Xa site" /translation="IEGR" CDS join(1259..1568,1898..2301) /codon_start=1 /label="mVenus" /note="mVenus" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" intron 1569..1897 /label="RBSC2i2" /note="RBSC2i2" CDS 2302..2313 /codon_start=1 /product="Factor Xa recognition and cleavage site" /note="Factor Xa site" /translation="IEGR" CDS 2326..2349 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /note="Strep-Tag II" /translation="WSHPQFEK" misc_feature 2353..2586 /label="3'UTR" /note="3'UTR" promoter 2599..2865 /label="HSP70Ap(1)" /note="HSP70Ap" promoter 2872..3092 /label="RBCS2p" /note="RBCS2p promoter" transit_peptide join(3093..3100,3246) /note="RBCS2ORF" intron 3101..3245 /label="RBCS2i(1)" /note="RBCS2i" CDS join(3253..3420,3566..3778) /codon_start=1 /note="ble" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPW GREFALRDPAGNCVHFVAEEQD" intron 3421..3565 /label="RBCS2_intron" /note="RBCS2_intron" misc_feature 3788..4026 /label="3'UTR(1)" /note="3'UTR" primer_bind complement(4086..4104) /label="M13_reverse_primer" /note="M13_reverse_primer" primer_bind complement(4103..4125) /label="M13_pUC_rev_primer" /note="M13_pUC_rev_primer" promoter complement(4139..4168) /label="lac_promoter" /note="lac_promoter" rep_origin complement(4477..5096) /direction=LEFT /label="pBR322ori" /note="pBR322ori" CDS complement(5245..6105) /codon_start=1 /label="Ampicillin" /note="Ampicillin" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6153..6180) /label="AmpRp" /note="AmpRp promoter" ORIGIN 1 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 61 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 121 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 181 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 241 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 301 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt 361 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct 421 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc 481 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt 541 ggagctccac cgcggtggcg gccgctctag agctgaggct tgacatgatt ggtgcgtatg 601 tttgtatgaa gctacaggac tgatttggcg ggctatgagg gcgggggaag ctctggaagg 661 gccgcgatgg ggcgcgcggc gtccagaagg cgccatacgg cccgctggcg gcacccatcc 721 ggtataaaag cccgcgaccc cgaacggtga cctccacttt cagcgacaaa cgagcactta 781 tacatacgcg actattctgc cgctatacat aaccactcag ctagcttaag atcccatcac 841 cggtgcatgc cgggcgcgcc agaaggagcg cagccaaacc aggatgatgt ttgatggggt 901 atttgagcac ttgcaaccct tatccggaag ccccctggcc cacaaaggct aggcgccaat 961 gcaagcagtt cgcatgcagc ccctggagcg gtgccctcct gataaaccgg ccagggggcc 1021 tatgttcttt acttttttac aagagaagtc actcaacatc ttaaaatggc caggtgagtc 1081 gacgagcaag cccggcggat caggcagcgt gcttgcagat ttgacttgca acgcccgcat 1141 tgtgtcgacg aaggcttttg gctcctctgt cgctgtctca agcagcatct aaccctgcgt 1201 cgccgtttcc atttgcagga tgcatatggg atccagatct gacgtcatcg agggcagggt 1261 gagcaagggc gaggagctgt tcaccggcgt ggtgcccatc ctggtggagc tggacggcga 1321 cgtgaacggc cacaagttca gcgtgagcgg cgagggcgag ggcgacgcca cctacggcaa 1381 gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctggt 1441 gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca 1501 cgacttcttc aagagcgcca tgcccgaggg ctacgtgcag gagcgcacca tcttcttcaa 1561 ggacgacggt gagcttgcgg ggttgcgagc aacactccag caacgaacag tgcccaagtc 1621 aggaatctgc agtcagcctg ggctttcggc ggctttttct tgggcaaaca gcttgcactc 1681 atgccagcgc ggcttgtcca gcctcacttg agctttccag ctgctaccag ccgggctata 1741 cgacagcgac agagccatag cgtggaatca cttatttggg ttgccgaagt agcggtcgga 1801 gcgtgagttc ttggtcaagc cgccccttat ccggttcctg tccgtgtctt tgtccctcgt 1861 tcacccttcg cggcaccctt catccccttg cttgcaggta actacaagac ccgcgccgag 1921 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag 1981 gaggacggca acatcctggg ccacaagctg gagtacaact acaacagcca caacgtgtac 2041 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc 2101 gaggacggcg gcgtgcagct ggccgaccac taccagcaga acacccccat cggcgacggc 2161 cccgtgctgc tgcccgacaa ccactacctg agctaccaga gcaagctgag caaggacccc 2221 aacgagaagc gcgaccacat ggtgctgctg gagttcgtga ccgccgccgg catcaccctg 2281 ggcatggacg agctgtacaa gatcgagggc agggatatcg aattctggag ccacccgcag 2341 ttcgagaagt aaccgctccg tgtaaatgga ggcgctcgtt gatctgagcc ttgccccctg 2401 acgaacggcg gtggatggaa gatactgctc tcaagtgctg aagcggtagc ttagctcccc 2461 gtttcgtgct gatcagtctt tttcaacacg taaaaagcgg aggagttttg caattttgtt 2521 ggttgtaacg atcctccgtt gattttggcc tctttctcca tgggcgggct gggcgtattt 2581 gaagcgacta gtacgcgtgc tgaggcttga catgattggt gcgtatgttt gtatgaagct 2641 acaggactga tttggcgggc tatgagggcg ggggaagctc tggaagggcc gcgatggggc 2701 gcgcggcgtc cagaaggcgc catacggccc gctggcggca cccatccggt ataaaagccc 2761 gcgaccccga acggtgacct ccactttcag cgacaaacga gcacttatac atacgcgact 2821 attctgccgc tatacataac cactcagcta gcttaagatc ccatccctag ggcatgccgg 2881 gcgcgccaga aggagcgcag ccaaaccagg atgatgtttg atggggtatt tgagcacttg 2941 caacccttat ccggaagccc cctggcccac aaaggctagg cgccaatgca agcagttcgc 3001 atgcagcccc tggagcggtg ccctcctgat aaaccggcca gggggcctat gttctttact 3061 tttttacaag agaagtcact caacatctta aaatggccag gtgagtcgac gagcaagccc 3121 ggcggatcag gcagcgtgct tgcagatttg acttgcaacg cccgcattgt gtcgacgaag 3181 gcttttggct cctctgtcgc tgtctcaagc agcatctaac cctgcgtcgc cgtttccatt 3241 tgcaggaagc ttatggccaa gctgaccagc gccgttccgg tgctcaccgc gcgcgacgtg 3301 gccggagcgg tcgagttctg gaccgaccgg ctcgggttca gccgggactt cgtggaggac 3361 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag 3421 gtgagtcgac gagcaagccc ggcggatcag gcagcgtgct tgcagatttg acttgcaacg 3481 cccgcattgt gtcgacgaag gcttttggct cctctgtcgc tgtctcaagc agcatctaac 3541 cctgcgtcgc cgtttccatt tgcaggacca ggtggtgccg gacaacaccc tggcctgggt 3601 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt 3661 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt 3721 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactaact 3781 cgagtgaccg ctccgtgtaa atggaggcgc tcgttgatct gagccttgcc ccctgacgaa 3841 cggcggtgga tggaagatac tgctctcaag tgctgaagcg gtagcttagc tccccgtttc 3901 gtgctgatca gtctttttca acacgtaaaa agcggaggag ttttgcaatt ttgttggttg 3961 taacgatcct ccgttgattt tggcctcttt ctccatgggc gggctgggcg tatttgaagc 4021 ggacccggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc 4081 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 4141 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 4201 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 4261 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 4321 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 4381 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 4441 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 4501 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 4561 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 4621 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 4681 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 4741 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 4801 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 4861 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 4921 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 4981 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 5041 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5101 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5161 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5221 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5281 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5341 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 5401 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 5461 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 5521 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 5581 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 5641 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 5701 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 5761 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 5821 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 5881 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 5941 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6001 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6061 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6121 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6181 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacactaaat 6241 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt 6301 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaaga //