LOCUS Exported 9729 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS BBa_J72114-BBa_J72100 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9729) AUTHORS Kittleson JT, DeLoache W, Cheng HY, Anderson JC TITLE Scalable plasmid transfer using engineered P1-based phagemids. JOURNAL ACS Synth Biol. 2012 Dec 21;1(12):583-9. doi: 10.1021/sb300054p. Epub 2012 Aug 30. PUBMED 23656280 REFERENCE 2 (bases 1 to 9729) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9729 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 77..3151 /codon_start=1 /gene="lacZ" /product="beta-galactosidase" /label=lacZ /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK" primer_bind complement(97..114) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(97..113) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(106..128) /label=M13/pUC Forward /note="In lacZ gene" terminator 3167..3261 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3282..3941) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 3856..3875 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" primer_bind complement(4202..4219) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4336..4881) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(4416..4435) /label=pBR322ori-F /note="pBR322 origin, forward primer" protein_bind 4961..4985 /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" terminator 8121..8148 /label=T7Te terminator /note="phage T7 early transcription terminator" terminator 8164..8235 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" promoter complement(8487..8771) /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind complement(8545..8564) /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" CDS 8798..9676 /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" ORIGIN 1 gaattcatga gatctgttaa tgctagctca gtcctaggga ctctgctagc ggatctattt 61 cactagggaa gccactatga ccatgattac ggattcactg gccgtcgttt tacaacgtcg 121 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc 181 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct 241 gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg aaagctggct 301 ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg 361 ttacgatgcg cccatctaca ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt 421 tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg aaagctggct 481 acaggaaggc cagacgcgaa ttatttttga tggcgttaac tcggcgtttc atctgtggtg 541 caacgggcgc tgggtcggtt acggccagga cagtcgtttg ccgtctgaat ttgacctgag 601 cgcattttta cgcgccggag aaaaccgcct cgcggtgatg gtgctgcgct ggagtgacgg 661 cagttatctg gaagatcagg atatgtggcg gatgagcggc attttccgtg acgtctcgtt 721 gctgcataaa ccgactacac aaatcagcga tttccatgtt gccactcgct ttaatgatga 781 tttcagccgc gctgtactgg aggctgaagt tcagatgtgc ggcgagttgc gtgactacct 841 acgggtaaca gtttctttat ggcagggtga aacgcaggtc gccagcggca ccgcgccttt 901 cggcggtgaa attatcgatg agcgtggtgg ttatgccgat cgcgtcacac tacgtctgaa 961 cgtcgaaaac ccgaaactgt ggagcgccga aatcccgaat ctctatcgtg cggtggttga 1021 actgcacacc gccgacggca cgctgattga agcagaagcc tgcgatgtcg gtttccgcga 1081 ggtgcggatt gaaaatggtc tgctgctgct gaacggcaag ccgttgctga ttcgaggcgt 1141 taaccgtcac gagcatcatc ctctgcatgg tcaggtcatg gatgagcaga cgatggtgca 1201 ggatatcctg ctgatgaagc agaacaactt taacgccgtg cgctgttcgc attatccgaa 1261 ccatccgctg tggtacacgc tgtgcgaccg ctacggcctg tatgtggtgg atgaagccaa 1321 tattgaaacc cacggcatgg tgccaatgaa tcgtctgacc gatgatccgc gctggctacc 1381 ggcgatgagc gaacgcgtaa cgcgaatggt gcagcgcgat cgtaatcacc cgagtgtgat 1441 catctggtcg ctggggaatg aatcaggcca cggcgctaat cacgacgcgc tgtatcgctg 1501 gatcaaatct gtcgatcctt cccgcccggt gcagtatgaa ggcggcggag ccgacaccac 1561 ggccaccgat attatttgcc cgatgtacgc gcgcgtggat gaagaccagc ccttcccggc 1621 tgtgccgaaa tggtccatca aaaaatggct ttcgctacct ggagagacgc gcccgctgat 1681 cctttgcgaa tacgcccacg cgatgggtaa cagtcttggc ggtttcgcta aatactggca 1741 ggcgtttcgt cagtatcccc gtttacaggg cggcttcgtc tgggactggg tggatcagtc 1801 gctgattaaa tatgatgaaa acggcaaccc gtggtcggct tacggcggtg attttggcga 1861 tacgccgaac gatcgccagt tctgtatgaa cggtctggtc tttgccgacc gcacgccgca 1921 tccagcgctg acggaagcaa aacaccagca gcagtttttc cagttccgtt tatccgggca 1981 aaccatcgaa gtgaccagcg aatacctgtt ccgtcatagc gataacgagc tcctgcactg 2041 gatggtggcg ctggatggta agccgctggc aagcggtgaa gtgcctctgg atgtcgctcc 2101 acaaggtaaa cagttgattg aactgcctga actaccgcag ccggagagcg ccgggcaact 2161 ctggctcaca gtacgcgtag tgcaaccgaa cgcgaccgca tggtcagaag ccgggcacat 2221 cagcgcctgg cagcagtggc gtctggcgga aaacctcagt gtgacgctcc ccgccgcgtc 2281 ccacgccatc ccgcatctga ccaccagcga aatggatttt tgcatcgagc tgggtaataa 2341 gcgttggcaa tttaaccgcc agtcaggctt tctttcacag atgtggattg gcgataaaaa 2401 acaactgctg acgccgctgc gcgatcagtt cacccgtgca ccgctggata acgacattgg 2461 cgtaagtgaa gcgacccgca ttgaccctaa cgcctgggtc gaacgctgga aggcggcggg 2521 ccattaccag gccgaagcag cgttgttgca gtgcacggca gatacacttg ctgatgcggt 2581 gctgattacg accgctcacg cgtggcagca tcaggggaaa accttattta tcagccggaa 2641 aacctaccgg attgatggta gtggtcaaat ggcgattacc gttgatgttg aagtggcgag 2701 cgatacaccg catccggcgc ggattggcct gaactgccag ctggcgcagg tagcagagcg 2761 ggtaaactgg ctcggattag ggccgcaaga aaactatccc gaccgcctta ctgccgcctg 2821 ttttgaccgc tgggatctgc cattgtcaga catgtatacc ccgtacgtct tcccgagcga 2881 aaacggtctg cgctgcggga cgcgcgaatt gaattatggc ccacaccagt ggcgcggcga 2941 cttccagttc aacatcagcc gctacagtca acagcaactg atggaaacca gccatcgcca 3001 tctgctgcac gcggaagaag gcacatggct gaatatcgac ggtttccata tggggattgg 3061 tggcgacgac tcctggagcc cgtcagtatc ggcggaattt cagctgagcg ccggtcgcta 3121 ccattaccag ttggtctggt gtcaaaaata aggatccgag gcttggattc tcaccaataa 3181 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta 3241 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac 3301 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg 3361 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg 3421 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg 3481 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg 3541 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc 3601 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa 3661 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata 3721 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac 3781 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg 3841 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg 3901 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct 3961 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag 4021 ttggaacctc ttacgtgccc gatcaaagat ccgcaccgcc ggacatcagc gctagcggag 4081 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca 4141 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt 4201 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg 4261 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc 4321 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg 4381 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 4441 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt 4501 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca 4561 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact 4621 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta 4681 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca 4741 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag 4801 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg 4861 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctaaggc 4921 ctcccctgat tctgtggata accgggatct gtaaggatca accactttgt acaagaaagc 4981 tgggtcgaat tgagatccga acggtttatt acgtacatca ggtaaaactg accgataagc 5041 cgctttcttt tgggtatagt gtcgtggaca gtcattcatc tttctgcccc tccaaaagta 5101 aaaacccgcc gaagcgggtt tttacgtaaa acaggtgaaa ctgaccgata agccgctttc 5161 ttttgggtat agtgtcgtgg acagtcattc atctttctgc ccctccaaaa gcaaaaaccc 5221 gccgaagcgg gtttttacgt aaaccaggtg aaactgaccg ataagccgct ttcttttggg 5281 tatagcgtcg tggacagtca ttcatctttc cgcccctcca aaagcaaaaa cccgccgaag 5341 cgggttttta cgtaaatcag gtgaaactga ccgataagcc gggttctgtc gtggacagtc 5401 attcatctag gccagcaatc gctcagatcc ttaatcatca tccccgtaca gtttgcgggt 5461 aacttcttcg tcttcctcct cgtcttcgtc cagatcctgt tcaagccatg cttcgtttga 5521 tatgccttcc gcatcgacat caccataacc acctgtatcg acgatatcgg caatttcttc 5581 tctacgatgc tcaatccaca atgcggcatc agcgcggcgg ctggcggccc gttctcgcgc 5641 gattttatcc agatcctcaa gtgatggagc gccagatgct gtttggtttt cctcatcatc 5701 ggtattggtc ttaggagcac gcagatcggc tttgatttgc tccagcatca ggggcggcac 5761 ttttcctcca tgcgcctcga tgaactcagc cgcttccagc actgaccagt tattttcacg 5821 ctttcgttcg tatgccagct taacaatgcc agcttgcccc atagataaag cgtgcttttc 5881 cgcctcccgg ctttcttttc gatagttatt ccggatgctg taaatggtgt tgatcaggct 5941 gcttatctgc gcggaacagc tgtttagcat gctcgcgata cggtattcag gcggagtacc 6001 ttcatcatcg tctttttgct gatcgcgcat ttcctgcacc aggcgaatac acgtatccct 6061 ggcgttctcc agcataagga gatgagaaag agacttttcc agaagagtgg tttccagaac 6121 atcggccccg gaccgacgca acatagcgcg cgcggccttc cgcgcttcaa cgttatctat 6181 caggtaatcg ccaacttcga actcaaagcg ttcaccatca tcatccaggg tgtcgcgttc 6241 caggcgatca cgtaaggtcc ggtgggcgcg ggtgatcacg tcatgatcat ctgaacgatc 6301 atttatgcgc ttattttggc gcttcgcatt ctcgactgcg gcactgacaa cggcattaac 6361 tctttgtttt tccgctattt cagccgcaat gtgatcacct gcatgttgat cattagagtg 6421 atcaatgatc atgcttttta gtggcttcct gactggctta tttggcttgc ggctgtccgt 6481 agtcctggtg tcttctttga aggcacggag ataacgacgt gcggtattag ggttaagatt 6541 aaactcggcg gcatactgtg cgatggtgta accaccatct cgcgccaggc gagcaaaatt 6601 cttcttgtga tcgtcccagg tcacttatgc ttcctttcga gatcccaaca atattcctac 6661 tcaatgaaca aatgactact cgtagaatcg gttaacacac cagattcaga tccttaccct 6721 ctgaatcctg ccggtatacc ccattgttcg ttatctttat ttttggctaa aaccgcatta 6781 agagcttcgt ttaccgtcat gcaatgcggt aggttatcga agtttgatat cccgccaata 6841 tcaggcgaac gcttgttttt caggtaagca tatttccgcg cagccgcctc tactttctgc 6901 ttgaactcat gtttttgagt gcgttttttg gataaccgca gattgtcagc ctttgctttt 6961 gccttagcga tccatgaagt caattttttg aggctggttg ttccggcacc gccggaaact 7021 gatctttttg tttttttaac ttgtgacttc ttattcttta ttgccacgtc atcctgacag 7081 ggggaggggg tatcattttg acatgggggt gtggataaaa aattaaataa agccaatgtc 7141 ttagcgagaa cagctttaac cttggttgcc gctgacgaaa tctttaattt gctttctatc 7201 agcgcatttt tggcttgttg tgcgaaggcc aaaaaggatg gtgtaaaccg gtacaggtta 7261 gcgcgacgtt cacggtgatc gccgataaca atctctacag acagaatacc tttgtttaca 7321 gcttcacgga atgcacgaac gacggttgat tggctataac cagtttctgc cgcgatcagg 7381 cggtgaggct tgtgaatgaa gtattcactg gttgttgccg cgagatttgc acattgcgac 7441 aggatatgcc cggcgctacg ggatagaccg gagtgtgtta caaagcaggc caattcatag 7501 ccagaaaaag taaaatcgct ttaagatccc cgagttctct taaaccaagg tttaggattg 7561 aaatgatgac gccggaaact tcttataaag cgtggaaaca gccacatcat agatgattgc 7621 aacctgctta cgggggatgc ccttctccag caatcgccgc atttgctgcc atgtttcttc 7681 ttggtattta ggccgacgcc cacctatacg accttctgcg cgagctgcat caagtccagc 7741 gcgtgtacgt tcaacgataa gctcacgttc catttctgcc agcgccccca ttacgtgaaa 7801 gaaaaagcgc cccattggtg tactggtgtc gatggagtca gtgagactcc ggaagttaat 7861 gcctctgtca cgcagctctt ccaccagcac aactaagtga cgcatgctgc gcccaagacg 7921 gtctaacttc catacgacca gggtatcacc tctggaaagt tacggagtac cttttttaac 7981 ccagggcgct cagccttttt gccgctcgcc ttgtcctcaa aaattagctc acatcctgcg 8041 ctttcaagag cgtttcgttg taaagcagtg ttttgttcat ttgttgatac gcgtacatag 8101 cctattagag atcctataaa cgcagaaagg cccacccgaa ggtgagccag tgtgactcta 8161 gtagagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc 8221 ctttcgtttt atttgatgcc tggagatcct tattgtgagt ctggctggtt agtggccatg 8281 agatattcga ttgtgtcagt gagatcatcc aggtcgtctt gggtgatgcg gtactcctga 8341 ttggatatct ttgagtagtg ttcagcaatg gcgcgggcag cgtcggtttc ggcggggtct 8401 acagataaag cgttagagca atgtctaacg tcgtcgatgg ttggtggaat gaaagccata 8461 attatgcctc actgtaagat ccgggtatgg agaaacagta gagagttgcg ataaaaagcg 8521 tcaggtaaga tccgctaatc ttatggataa aaatgctatg gcatagcaaa gtgtgacgcc 8581 gtgcaaataa tcaatgtgga cttttctgcc gtgattatag acacttttgt tacgcgtttt 8641 tgtcatggct ttggtcccgc tttgttacag aatgctttta ataagcgggg ttaccggttt 8701 ggttagcgag aagagccagt aaaagacgca gtgacggcaa tgtctgatgc aatatggaca 8761 attggtttct tctctgaatg gcgggagtat gaaaagtatg gctgaagcgc aaaatgatcc 8821 cctgctgccg ggatactcgt ttaatgccca tctggtggcg ggtttaacgc cgattgaggc 8881 caacggttat ctcgattttt ttatcgaccg accgctggga atgaaaggtt atattctcaa 8941 tctcaccatt cgcggtcagg gggtggtgaa aaatcaggga cgagaatttg tttgccgacc 9001 gggtgatatt ttgctgttcc cgccaggaga gattcatcac tacggtcgtc atccggaggc 9061 tcgcgaatgg tatcaccagt gggtttactt tcgtccgcgc gcctactggc atgaatggct 9121 taactggccg tcaatatttg ccaatacggg gttctttcgc ccggatgaag cgcaccagcc 9181 gcatttcagc gacctgtttg ggcaaatcat taacgccggg caaggggaag ggcgctattc 9241 ggagctgctg gcgataaatc tgcttgagca attgttactg cggcgcatgg aagcgattaa 9301 cgagtcgctc catccaccga tggataatcg ggtacgcgag gcttgtcagt acatcagcga 9361 tcacctggca gacagcaatt ttgatatcgc cagcgtcgca cagcatgttt gcttgtcgcc 9421 gtcgcgtctg tcacatcttt tccgccagca gttagggatt agcgtcttaa gctggcgcga 9481 ggaccaacgt atcagccagg cgaagctgct tttgagcacc acccggatgc ctatcgccac 9541 cgtcggtcgc aatgttggtt ttgacgatca actctatttc tcgcgggtat ttaaaaaatg 9601 caccggggcc agcccgagcg agttccgtgc cggttgtgaa gaaaaagtga atgatgtagc 9661 cgtcaagttg tcataattgg taacgaatca gacaattgac ggctcgatgg agtagcatag 9721 agatctcat //