LOCUS       sequence-452443-        5116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Hybrid Reporter Plasmid (sfGFP), pT7-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5116)
  AUTHORS   Baumer ZT, Newton MS, Lofstrand L, Carpio Paucar GN, Farny NG, 
            Whitehead TA
  TITLE     Engineered Stop and Go T7 RNA Polymerases.
  JOURNAL   ACS Synth Biol. 2024 Dec 20;13(12):4165-4174. doi: 
            10.1021/acssynbio.4c00627. Epub 2024 Nov 28.
  PUBMED    39610115
REFERENCE   2  (bases 1 to 5116)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.4c00627"; journalName: "ACS Synth Biol"; date: 
            "2024-12-20- 20"; volume: "13"; issue: "12"; pages: "4165-4174"
COMMENT     Sequence Label: pZB604_pT7-sfGFP
FEATURES             Location/Qualifiers
     source          1..5116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(578..672)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(703..1497)
                     /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"
     primer_bind     complement(814..833)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1424..1443
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      1652..1738
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1830..1857
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1935..1953
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     1996..2015
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1996..2014
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2042..2755
                     /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
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2767..2838
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2854..2881
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2890..3345)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3027..3048)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3239..3258
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      3358..3444
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3982..4932)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      join(4980..5116,1..86)
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa gataaaaaga
       61 atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta ttacaaaagg
      121 atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa aggcttaagt
      181 agcaccctcg caagctcggg caaatcgctg aatattcctt ttgtctccga ccatcaggca
      241 cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc tggcagtgaa
      301 tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac tacccataat
      361 acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc
      421 tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct gaccacttcg
      481 gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca gcggtatcat
      541 caacaggctt acccgtctta ctgtccctag tgcttggatt ctcaccaata aaaaacgccc
      601 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
      661 tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag
      721 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
      781 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
      841 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
      901 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
      961 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     1021 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     1081 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     1141 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     1201 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     1261 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
     1321 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     1381 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     1441 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     1501 aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg
     1561 cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc
     1621 agctggcaat tccgacgtca ccggtaggca tcaaataaaa cgaaaggctc agtcgaaaga
     1681 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     1741 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     1801 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     1861 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1921 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaaacg cgtgtacacc
     1981 cgggcggccg caaattaata cgactcacta tagggagatt agctaagagg agaaatacta
     2041 gatgagcaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg
     2101 tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg
     2161 taaactgacg ctgaagttca tctgtactac tggtaaactg ccggttcctt ggccgactct
     2221 ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca
     2281 gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt
     2341 taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt
     2401 aaaccgcatt gagctgaaag gcattgactt taaagaggac ggcaatatcc tgggccataa
     2461 gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg
     2521 cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga
     2581 tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta
     2641 tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct
     2701 gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatgacc
     2761 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     2821 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     2881 gctcgagcta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2941 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3001 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3061 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3121 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3181 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3241 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3301 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtctcta gaggcatcaa
     3361 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3421 aacgctctcc tgagtaggac aaatccgccg ccctagacct agggtacggg ttttgctgcc
     3481 cgcaaacggg ctgttctggt gttgctagtt tgttatcaga atcgcagatc cggcttcagc
     3541 cggtttgccg gctgaaagcg ctatttcttc cagaattgcc atgatttttt ccccacggga
     3601 ggcgtcactg gctcccgtgt tgtcggcagc tttgattcga taagcagcat cgcctgtttc
     3661 aggctgtcta tgtgtgactg ttgagctgta acaagttgtc tcaggtgttc aatttcatgt
     3721 tctagttgct ttgttttact ggtttcacct gttctattag gtgttacatg ctgttcatct
     3781 gttacattgt cgatctgttc atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc
     3841 tctgatgtat ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt
     3901 gatatgtaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata
     3961 gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt
     4021 ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatact tactttgcat
     4081 gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg
     4141 tagtgttttt cttagtccgt tatgtaggta ggaatctgat gtaatggttg ttggtatttt
     4201 gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc
     4261 tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt
     4321 catattgctg taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct
     4381 tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat
     4441 ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat
     4501 cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt
     4561 ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct
     4621 tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct
     4681 gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt
     4741 ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc
     4801 tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt
     4861 ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac
     4921 taattcagac atacatctca attggtctag gtgattttaa tcactatacc aattgagatg
     4981 ggctagtcaa tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg
     5041 ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc
     5101 tttgtgtgtt tttttt
//
