LOCUS Exported 4367 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION PT7 GFP reporter. ACCESSION . VERSION . KEYWORDS pTHSSd_8 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4367) AUTHORS Segall-Shapiro TH, Meyer AJ, Ellington AD, Sontag ED, Voigt CA TITLE A 'resource allocator' for transcription based on a highly fragmented T7 RNA polymerase. JOURNAL Mol Syst Biol. 2014 Jul 30;10(7):742. PUBMED 25080493 REFERENCE 2 (bases 1 to 4367) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4367 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 59..772 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(89..117) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 712..732 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 796..843 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(1418..2368) /codon_start=1 /gene="rep101" /product="RepA protein needed for replication with the pSC101 origin" /label=Rep101 /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI" rep_origin 2416..2638 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" terminator 3130..3224 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3255..4115) /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 3878..3897 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" primer_bind 4329..4347 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 taatacgact cactataggg agaggatcct gtcaatttcc gcgatagagg aggtaaagat 61 gagcaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga 121 tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa ggtgatgcta caaacggaaa 181 actcaccctt aaatttattt gcactactgg aaaactacct gttccgtggc caacacttgt 241 cactactctg acctatggtg ttcaatgctt ttcccgttat ccggatcaca tgaaacggca 301 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaacgcacta tatctttcaa 361 agatgacggg acctacaaga cgcgtgctga agtcaagttt gaaggtgata cccttgttaa 421 tcgtatcgag ttaaagggta ttgattttaa agaagatgga aacattcttg gacacaaact 481 cgagtacaac tttaactcac acaatgtata catcacggca gacaaacaaa agaatggaat 541 caaagctaac ttcaaaattc gccacaacgt tgaagatggt tccgttcaac tagcagacca 601 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct 661 gtcgacacaa tctgtccttt cgaaagatcc caacgaaaag cgtgaccaca tggtccttct 721 tgagtttgta actgctgctg ggattacaca tggcatggat gagctctaca aataataagg 781 tgcttaataa ggatcctagc ataacccctt ggggcctcta aacgggtctt gaggggtttt 841 ttgctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgc cctagaccta 901 gggtacgggt tttgctgccc gcaaacgggc tgttctggtg ttgctagttt gttatcagaa 961 tcgcagatcc ggcttcaggt ttgccggctg aaagcgctat ttcttccaga attgccatga 1021 ttttttcccc acgggaggcg tcactggctc ccgtgttgtc ggcagctttg attcgataag 1081 cagcatcgcc tgtttcaggc tgtctatgtg tgactgttga gctgtaacaa gttgtctcag 1141 gtgttcaatt tcatgttcta gttgctttgt tttactggtt tcacctgttc tattaggtgt 1201 tacatgctgt tcatctgtta cattgtcgat ctgttcatgg tgaacagctt taaatgcacc 1261 aaaaactcgt aaaagctctg atgtatctat cttttttaca ccgttttcat ctgtgcatat 1321 ggacagtttt ccctttgata tctaacggtg aacagttgtt ctacttttgt ttgttagtct 1381 tgatgcttca ctgatagata caagagccat aagaacctca gatccttccg tatttagcca 1441 gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa ccattgagat 1501 catgcttact ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg 1561 cagttaaagc atcgtgtagt gtttttctta gtccgttacg taggtaggaa tctgatgtaa 1621 tggttgttgg tattttgtca ccattcattt ttatctggtt gttctcaagt tcggttacga 1681 gatccatttg tctatctagt tcaacttgga aaatcaacgt atcagtcggg cggcctcgct 1741 tatcaaccac caatttcata ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa 1801 cccattggtt aagcctttta aactcatggt agttattttc aagcattaac atgaacttaa 1861 attcatcaag gctaatctct atatttgcct tgtgagtttt cttttgtgtt agttctttta 1921 ataaccactc ataaatcctc atagagtatt tgttttcaaa agacttaaca tgttccagat 1981 tatattttat gaattttttt aactggaaaa gataaggcaa tatctcttca ctaaaaacta 2041 attctaattt ttcgcttgag aacttggcat agtttgtcca ctggaaaatc tcaaagcctt 2101 taaccaaagg attcctgatt tccacagttc tcgtcatcag ctctctggtt gctttagcta 2161 atacaccata agcattttcc ctactgatgt tcatcatctg agcgtattgg ttataagtga 2221 acgataccgt ccgttctttc cttgtagggt tttcaatcgt ggggttgagt agtgccacac 2281 agcataaaat tagcttggtt tcatgctccg ttaagtcata gcgactaatc gctagttcat 2341 ttgctttgaa aacaactaat tcagacatac atctcaattg gtctaggtga ttttaatcac 2401 tataccaatt gagatgggct agtcaatgat aattactagt ccttttcctt tgagttgtgg 2461 gtatctgtaa attctgctag acctttgctg gaaaacttgt aaattctgct agaccctctg 2521 taaattccgc tagacctttg tgtgtttttt ttgtttatat tcaagtggtt ataatttata 2581 gaataaagaa agaataaaaa aagataaaaa gaatagatcc cagccctgtg tataactcac 2641 tactttagtc agttccgcag tattacaaaa ggatgtcgca aacgctgttt gctcctctac 2701 aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg ggcaaatcgc 2761 tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt tcgtgacatt 2821 cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta caggcgcctt 2881 ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac gggcttctca 2941 gggcgtttta tggcgggtct gctatgtggt gctatctgac tttttgctgt tcagcagttc 3001 ctgccctctg attttccagt ctgaccactt cggattatcc cgtgacaggt cattcagact 3061 ggctaatgca cccagtaagg cagcggtatc atcaacaggc ttacccgtct tactgtccct 3121 agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc 3181 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctctcgaa 3241 ccccagagtc ccgcttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 3301 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 3361 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 3421 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 3481 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 3541 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 3601 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 3661 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 3721 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 3781 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 3841 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 3901 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 3961 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 4021 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 4081 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 4141 agcatttatc gcgaaacgat cctcatcctg tctcttgatc agatcttgat cccctgcgcc 4201 atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac 4261 cagagggcgc cccagctggc aattccgacg tctaagaaac cattattatc atgacattaa 4321 cctataaaaa taggcgtatc acgaggccct ttcgtcttca cctcgag //