LOCUS Exported 6335 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS J72008 bacterial strain with plasmid SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6335) AUTHORS Anderson JC, Dueber JE, Leguia M, Wu GC, Goler JA, Arkin AP, Keasling JD TITLE BglBricks: A flexible standard for biological part assembly. JOURNAL J Biol Eng. 2010 . 4(1):1. PUBMED 20205762 REFERENCE 2 (bases 1 to 6335) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6335 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(62..775) /codon_start=1 /gene="cIts" /product="temperature-sensitive variant of the phage lambda repressor" /label=-lambda repressor (ts) /note="thermosensitivity is conferred by the A67T mutation" /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV VGKVIASQWPEETFG" RBS 864..872 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" CDS 1009..2217 /codon_start=1 /gene="int" /product="integrase from phage phi-80" /label=phage phi-80 integrase /translation="MCTHSMITDTKLRKALGKKRDDIEIISDSHGLNARISQAGKISFF YRYRWAGKAVKLNVGDYPAMSITQARERRQQFRNWLTEGLDPREQVKLDKQTRQEAMSV AEAFNYWIERHCIANGLVKVDYYRQVFEKHIAEPMKNVKVDNTAKMHWINVFDSIESRV MAHYMLSLCKRAFRFCVNRSVIASNPLEGLLPSDVGQKPKKRTRRMDDDDLRKIYQWLK SHMSIESVFLVKFIMLTGCRTAEIRLSERSWFRLDDNEWVVPAGSYKTRVHIRRGLSDA AVNLVRNHLKKINTNHLVTSQRKIDGGIKDSPVHSPVASNYARSIWNGTGMAEWSLHDM RRTIATNLSELGCPPHVIEKLLGHQMVGVMAHYNLHDYIDDQKHWLRVWQSHLEEIIGE PFS" CDS complement(3502..4452) /codon_start=1 /gene="rep101" /product="RepA protein needed for replication with the pSC101 origin" /label=Rep101 /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI" rep_origin 4500..4722 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" CDS complement(5344..6204) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5967..5986 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6205..6309) /gene="bla" /label=AmpR promoter ORIGIN 1 catcgatgaa gattcttgct caattgttat cagctatgcg ccgaccagaa caccttgccg 61 atcagccaaa cgtctcttca ggccactgac tagcgataac tttccccaca acggaacaac 121 tctcattgca tgggatcatt gggtactgtg ggtttagtgg ttgtaaaaac acctgaccgc 181 tatccctgat cagtttcttg aaggtaaact catcaccccc aagtctggct atgcagaaat 241 cacctggctc aacagcctgc tcagggtcaa cgagaattaa cattccgtca ggaaagcttg 301 gcttggagcc tgttggtgcg gtcatggaat taccttcaac ctcaagccag aatgcagaat 361 cactggcttt tttggttgtg cttacccatc tctccgcatc acctttggta aaggttctaa 421 gcttaggtga gaacatccct gcctgaacat gagaaaaaac agggtactca tactcacttc 481 taagtgacgg ctgcatacta accgcttcat acatctcgta gatttctctg gcgattgaag 541 ggctaaattc ttcaacgcta actttgagaa tttttgtaag caatgcggcg ttataagcat 601 ttaatgcatt gatgccatta aataaagcac caacgcctga ctgccccatc cccatcttgt 661 ctgcgacaga ttcctgggat aagccaagtt catttttctt tttttcataa attgctttaa 721 ggcgacgtgc gtcctcaagc tgctcttgtg ttaatggttt cttttttgtg ctcatacgtt 781 aaatctatca ccgcaaggga taaatatcta acaccgtgcg tgttgactat tttacctctg 841 gcggtgataa tggttgcatg tactaaggag gttgtatgaa ttcgctagca acccttatcc 901 agcaggcttt caaggatgta aaccataaca ctctgcgaac tagtgttaca ttgcgtgtag 961 ctttgagtgg gcaactttgt gtacactttt gtgtacccaa aaacaaaaat gtgtacccat 1021 tcaatgatca ccgacacaaa gctcaggaag gcgctcggca agaaaagaga tgatatcgag 1081 attatttctg attcgcacgg gctcaacgcc agaatcagcc aggccggaaa aatatcattt 1141 ttctatcggt atcgctgggc cggtaaagcg gtaaaactca atgttggtga ttatcctgca 1201 atgagtatca cccaggcaag agagcgtcgc caacaattca gaaactggtt aactgaggga 1261 ctggatccgc gagagcaggt gaagctggat aagcagaccc gacaggaagc gatgtccgtt 1321 gccgaagcgt tcaattactg gattgaaagg cactgtatcg ctaacgggct agttaaagtc 1381 gattactatc gccaggtgtt tgagaaacat atcgccgaac cgatgaagaa tgtcaaagtc 1441 gataacacag cgaaaatgca ctggatcaac gtcttcgatt ctatagaaag cagggtgatg 1501 gctcattaca tgctttcgct gtgcaaacgg gcgtttaggt tctgcgttaa cagaagtgtg 1561 atcgcctcaa acccgctcga gggattactg ccatctgatg tcgggcaaaa gcctaaaaag 1621 agaactcgca ggatggacga tgacgatctg cgcaaaatct atcagtggtt gaaaagccat 1681 atgtcgatag agtccgtttt cctggtgaaa tttattatgc ttaccggatg ccgtacggct 1741 gagattcgac ttagtgagag atcatggttt cgattggatg ataatgagtg ggtcgtgcct 1801 gcgggcagtt ataaaactcg ggtacatatt agaaggggac tctcagacgc cgccgttaac 1861 ctggtcagaa atcacctcaa gaaaataaac accaatcacc tggtgacttc acaacgtaaa 1921 attgatggcg ggatcaaaga ttcgcccgtt cattcacctg tggcatccaa ttacgcccgt 1981 tctatttgga atggaacagg tatggcagag tggtcgcttc atgatatgag gcggacgata 2041 gccacaaatc tctctgagtt aggttgcccg ccgcacgtaa ttgaaaagct gctcgggcat 2101 cagatggtgg gggttatggc gcattacaac cttcatgact atatcgatga tcagaaacac 2161 tggctccgcg tttggcagag ccatcttgaa gagatcatcg gagagccctt cagttaattt 2221 atcttctttt tatcctccca ctctttgatt gccatgggta tggacagttt tccctttgtc 2281 tagacatgag tggatagtac gttgctaaaa catgagataa aaattgactc tcatgttatt 2341 ggcgttaaga tatacagaat gatgaggttt ttttatgaga ctcaaggtca tgatggacgt 2401 gaacaaaaaa acgaaaattc gccaccgaaa cgagctaaat cacaccctgg ctcaacttcc 2461 tttgcccgca aagcgagtga tgtatatggc gcttgctccc attgatagca aagaacctct 2521 tgaacgaggg cgagttttca aaattagggc tgaagacctt gcagcgctcg ccaaaatcac 2581 cccatcgctt gcttatcgac aattaaaaga gggtggtaaa ttacttggtg ccagcaaaat 2641 ttcgctaaga ggggatgata tcattgcttt agctaaagag cttaacctgc tctttactgc 2701 taaaaactcc cctgaagagt tagatcttaa cattattgag tggatagctt attcaaatga 2761 tgaaggatac ttgtctttaa aattcaccag aaccatagaa ccatatatct ctagccttat 2821 tgggaaaaaa aataaattca caacgcaatt gttaacggca agcttacgct taagtagcca 2881 gtattcatct tctctttatc aacttatcag gaagcattac tctaatttta agaagaaaaa 2941 ttattttatt atttccgttg atgagttaaa ggaagagtta acagcttata cttttgataa 3001 agatggaaat attgagtaca aataccctga ctttcctatt tttaaaaggg atgtgttaaa 3061 taaagccatt gctgaaatta aaaagaaaac agaaatatcg tttgttggct tcactgttca 3121 tgaaaaagaa ggaagaaaaa ttagtaagct gaagttcgaa tttgtcgttg atgaagatga 3181 attttctggc gataaagatg atgaagcttt ttttatgaat ttatctgaag ctgatgcagc 3241 ttttctcaag gtatttaatg aaaccgtacc tcccaaaaaa gctaaggggt gatatatggc 3301 taaaatttac gatttccctc aaggagccga acgccgcagg atgcaccgca aaatccagtg 3361 gaacaacgct gtaaaattat ctaaaaatgg ctggagtaag ccagaggtta aacgcagatc 3421 tatatgtaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata 3481 gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt 3541 ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatact tactttgcat 3601 gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg 3661 tagtgttttt cttagtccgt tacgtaggta ggaatctgat gtaatggttg ttggtatttt 3721 gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc 3781 tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt 3841 catattgctg taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct 3901 tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat 3961 ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat 4021 cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt 4081 ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct 4141 tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct 4201 gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt 4261 ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc 4321 tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt 4381 ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac 4441 taattcagac atacatctca attggtctag gtgattttaa tcactatacc aattgagatg 4501 ggctagtcaa tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg 4561 ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc 4621 tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata 4681 aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc 4741 gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa 4801 ccctaaaggc ttaagtagca ccctcgcaag ctcggttgcg gccgcaatcg ggcaaatcgc 4861 tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt tcgtgacatt 4921 cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta caggcgcctt 4981 ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac gggcttctca 5041 gggcgtttta tggcgggtct gctatgtggt gctatctgac tttttgctgt tcagcagttc 5101 ctgccctctg attttccagt ctgaccactt cggattatcc cgtgacaggt cattcagact 5161 ggctaatgca cccagtaagg cagcggtatc atcaacgggg tctgacgctc agtggaacga 5221 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 5281 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 5341 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 5401 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 5461 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 5521 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 5581 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 5641 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 5701 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 5761 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 5821 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 5881 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 5941 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 6001 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 6061 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 6121 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 6181 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 6241 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 6301 ggttccgcgc acatttcccc gaaaagtgcc acctg //