LOCUS Exported 5515 bp DNA circular SYN 30-NOV-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 5515) AUTHORS jeremygarb TITLE Direct Submission JOURNAL Exported Nov 30, 2021 from SnapGene 5.3.2 https://www.snapgene.com FEATURES Location/Qualifiers source 1..5515 /organism="recombinant plasmid" /mol_type="other DNA" CDS complement(7..885) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 912..1196 /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)" CDS 1246..1959 /codon_start=1 /gene="sfGFP(Sp)" /label=sfGFP(Sp) /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" misc_signal 1246..1248 /label=START CDS 1978..2067 /codon_start=1 /product="two Strep-Tag(R)II moieties connected by a linker for enhanced binding to Strep-Tactin(R), an engineered form of streptavidin (Schmidt et al., 2013)" /label=Twin-Strep-tag /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK" misc_feature 2068..2070 terminator 2103..2174 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2190..2217 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(2748..3698) /codon_start=1 /gene="rep101" /product="RepA protein needed for replication with the pSC101 origin" /label=Rep101 /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI" terminator 4460..4554 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(4585..5379) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" ORIGIN 1 gacgtcttat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa 61 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc 121 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt 181 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg 241 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc 301 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc 361 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag 421 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg 481 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt 541 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac 601 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg 661 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac 721 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat 781 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt 841 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc 901 gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt 961 ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca 1021 aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa 1081 agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat 1141 aagattagcg gattctacct gacgcttttt atcgcaactc tctactgttt ctccataccc 1201 gtttttttgg gaattcaaaa gatcttttaa gaaggagata tacatatgtc aaaaggagaa 1261 gaacttttta caggtgtagt acctatcttg gttgaattgg atggtgatgt taacggtcac 1321 aaattttctg tacgtggtga aggtgaaggt gatgcaacta acggtaaatt gacacttaaa 1381 ttcatttgta caactggaaa acttcctgtt ccttggccta ctcttgttac aacattgaca 1441 tatggagtac aatgtttttc acgttatcct gatcatatga aacgtcacga tttttttaaa 1501 tctgctatgc cagaaggtta tgtacaagaa cgtacaattt catttaaaga tgacggaaca 1561 tataaaacac gtgctgaagt aaaattcgaa ggtgacactc ttgttaatcg tatcgaattg 1621 aaaggaatcg atttcaaaga agatggtaac attttgggac acaaacttga atacaacttc 1681 aactctcata atgtttatat cacagctgac aaacaaaaaa acggtattaa agctaatttt 1741 aaaattcgtc acaatgttga agatggatct gttcaattgg ctgatcatta tcaacaaaat 1801 acaccaatcg gagacggacc agtattgctt ccagataacc actacctttc tactcaatca 1861 gttctttcaa aagatcctaa cgaaaaacgt gaccatatgg tacttcttga atttgttaca 1921 gcagcaggta tcactcacgg tatggacgaa ctttataaag ttgacacagg gaacggctca 1981 gcttggtcgc acccgcagtt tgaaaagggt ggaggttcgg gcggtgggag cggcggcagt 2041 gcgtggtcgc atcctcagtt tgaaaagtaa ggatccaaac tcgagtaagg atctccaggc 2101 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt 2161 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt 2221 atacctaggg tacgggtttt gctgcccgca aacgggctgt tctggtgttg ctagtttgtt 2281 atcagaatcg cagatccggc ttcagccggt ttgccggctg aaagcgctat ttcttccaga 2341 attgccatga ttttttcccc acgggaggcg tcactggctc ccgtgttgtc ggcagctttg 2401 attcgataag cagcatcgcc tgtttcaggc tgtctatgtg tgactgttga gctgtaacaa 2461 gttgtctcag gtgttcaatt tcatgttcta gttgctttgt tttactggtt tcacctgttc 2521 tattaggtgt tacatgctgt tcatctgtta cattgtcgat ctgttcatgg tgaacagctt 2581 tgaatgcacc aaaaactcgt aaaagctctg atgtatctat cttttttaca ccgttttcat 2641 ctgtgcatat ggacagtttt ccctttgata tgtaacggtg aacagttgtt ctacttttgt 2701 ttgttagtct tgatgcttca ctgatagata caagagccat aagaacctca gatccttccg 2761 tatttagcca gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa 2821 ccattgagat catacttact ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc 2881 tgaatttttg cagttaaagc atcgtgtagt gtttttctta gtccgttatg taggtaggaa 2941 tctgatgtaa tggttgttgg tattttgtca ccattcattt ttatctggtt gttctcaagt 3001 tcggttacga gatccatttg tctatctagt tcaacttgga aaatcaacgt atcagtcggg 3061 cggcctcgct tatcaaccac caatttcata ttgctgtaag tgtttaaatc tttacttatt 3121 ggtttcaaaa cccattggtt aagcctttta aactcatggt agttattttc aagcattaac 3181 atgaacttaa attcatcaag gctaatctct atatttgcct tgtgagtttt cttttgtgtt 3241 agttctttta ataaccactc ataaatcctc atagagtatt tgttttcaaa agacttaaca 3301 tgttccagat tatattttat gaattttttt aactggaaaa gataaggcaa tatctcttca 3361 ctaaaaacta attctaattt ttcgcttgag aacttggcat agtttgtcca ctggaaaatc 3421 tcaaagcctt taaccaaagg attcctgatt tccacagttc tcgtcatcag ctctctggtt 3481 gctttagcta atacaccata agcattttcc ctactgatgt tcatcatctg agcgtattgg 3541 ttataagtga acgataccgt ccgttctttc cttgtagggt tttcaatcgt ggggttgagt 3601 agtgccacac agcataaaat tagcttggtt tcatgctccg ttaagtcata gcgactaatc 3661 gctagttcat ttgctttgaa aacaactaat tcagacatac atctcaattg gtctaggtga 3721 ttttaatcac tataccaatt gagatgggct agtcaatgat aattactagt ccttttcccg 3781 ggtgatctgg gtatctgtaa attctgctag acctttgctg gaaaacttgt aaattctgct 3841 agaccctctg taaattccgc tagacctttg tgtgtttttt ttgtttatat tcaagtggtt 3901 ataatttata gaataaagaa agaataaaaa aagataaaaa gaatagatcc cagccctgtg 3961 tataactcac tactttagtc agttccgcag tattacaaaa ggatgtcgca aacgctgttt 4021 gctcctctac aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg 4081 ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt 4141 tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta 4201 caggcgcctt ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac 4261 gggcttctca gggcgtttta tggcgggtct gctatgtggt gctatctgac tttttgctgt 4321 tcagcagttc ctgccctctg attttccagt ctgaccactt cggattatcc cgtgacaggt 4381 cattcagact ggctaatgca cccagtaagg cagcggtatc atcaacaggc ttacccgtct 4441 tactgtccct agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc 4501 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg 4561 agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat 4621 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc 4681 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac 4741 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg 4801 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag 4861 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc 4921 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc 4981 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga 5041 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa 5101 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc 5161 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga 5221 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc 5281 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc 5341 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt 5401 ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca 5461 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attcc //