LOCUS       sequence-452522-        5807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Hybrid Reporter Plasmid, pT7-HIS3/URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5807)
  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 5807)
  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: pZB260_pT7-STR-H3U3
FEATURES             Location/Qualifiers
     source          1..5807
                     /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"
     primer_bind     2028..2050
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2047..2063
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2059..2721
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     CDS             2769..3572
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(3581..4036)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3718..3739)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3930..3949
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      4049..4135
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4673..5623)
                     /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(5671..5807,1..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 tagggagatg tcgaccgagc ggataacaat
     2041 ttcacacagg aaacagctat gacagagcag aaagccctag taaagcgtat tacaaatgaa
     2101 accaagattc agattgcgat ctctttaaag ggtggtcccc tagcgataga gcactcgatc
     2161 ttcccagaaa aagaggcaga agcagtagca gaacaggcca cacaatcgca agtgattaac
     2221 gtccacacag gtatagggtt tctggaccat atgatacatg ctctggccaa gcattccggc
     2281 tggtcgctaa tcgttgagtg cattggtgac ttacacatag acgaccatca caccactgaa
     2341 gactgcggga ttgctctcgg tcaagctttt aaagaggccc taggggccgt gcgtggagta
     2401 aaaaggtttg gatcaggatt tgcgcctttg gatgaggcac tttccagagc ggtggtagat
     2461 ctttcgaaca ggccgtacgc agttgtcgaa cttggtttgc aaagggagaa agtaggagat
     2521 ctctcttgcg agatgatccc gcattttctt gaaagctttg cagaggctag cagaattacc
     2581 ctccacgttg attgtctgcg aggcaagaat gatcatcacc gtagtgagag tgcgttcaag
     2641 gctcttgcgg ttgccataag agaagccacc tcgcccaatg gtaccaacga tgttccctcc
     2701 accaaaggtg ttcttatgta gtgacaccga ttatttaaag ctgcagggat cccacacagg
     2761 aaacagctat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg
     2821 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg
     2881 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac
     2941 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa
     3001 aggcattatc cgccaagtac aattttttac tcttcgaaga cagaaaattt gctgacattg
     3061 gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca
     3121 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag
     3181 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct
     3241 ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt
     3301 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt
     3361 tgattatgac acccggtgtg ggtttagatg acaagggaga cgcattgggt caacagtata
     3421 gaaccgtgga tgatgtggtc tctacaggat ctgacattat tattgttgga agaggactat
     3481 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag
     3541 catatttgag aagatgcggc cagcaaaact aactcgagct aaattgtaag cgttaatatt
     3601 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3661 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3721 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3781 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3841 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3901 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3961 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4021 ccgctacagg gcgcgtctct agaggcatca aataaaacga aaggctcagt cgaaagactg
     4081 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     4141 gccctagacc tagggtacgg gttttgctgc ccgcaaacgg gctgttctgg tgttgctagt
     4201 ttgttatcag aatcgcagat ccggcttcag ccggtttgcc ggctgaaagc gctatttctt
     4261 ccagaattgc catgattttt tccccacggg aggcgtcact ggctcccgtg ttgtcggcag
     4321 ctttgattcg ataagcagca tcgcctgttt caggctgtct atgtgtgact gttgagctgt
     4381 aacaagttgt ctcaggtgtt caatttcatg ttctagttgc tttgttttac tggtttcacc
     4441 tgttctatta ggtgttacat gctgttcatc tgttacattg tcgatctgtt catggtgaac
     4501 agctttgaat gcaccaaaaa ctcgtaaaag ctctgatgta tctatctttt ttacaccgtt
     4561 ttcatctgtg catatggaca gttttccctt tgatatgtaa cggtgaacag ttgttctact
     4621 tttgtttgtt agtcttgatg cttcactgat agatacaaga gccataagaa cctcagatcc
     4681 ttccgtattt agccagtatg ttctctagtg tggttcgttg tttttgcgtg agccatgaga
     4741 acgaaccatt gagatcatac ttactttgca tgtcactcaa aaattttgcc tcaaaactgg
     4801 tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg ttatgtaggt
     4861 aggaatctga tgtaatggtt gttggtattt tgtcaccatt catttttatc tggttgttct
     4921 caagttcggt tacgagatcc atttgtctat ctagttcaac ttggaaaatc aacgtatcag
     4981 tcgggcggcc tcgcttatca accaccaatt tcatattgct gtaagtgttt aaatctttac
     5041 ttattggttt caaaacccat tggttaagcc ttttaaactc atggtagtta ttttcaagca
     5101 ttaacatgaa cttaaattca tcaaggctaa tctctatatt tgccttgtga gttttctttt
     5161 gtgttagttc ttttaataac cactcataaa tcctcataga gtatttgttt tcaaaagact
     5221 taacatgttc cagattatat tttatgaatt tttttaactg gaaaagataa ggcaatatct
     5281 cttcactaaa aactaattct aatttttcgc ttgagaactt ggcatagttt gtccactgga
     5341 aaatctcaaa gcctttaacc aaaggattcc tgatttccac agttctcgtc atcagctctc
     5401 tggttgcttt agctaataca ccataagcat tttccctact gatgttcatc atctgagcgt
     5461 attggttata agtgaacgat accgtccgtt ctttccttgt agggttttca atcgtggggt
     5521 tgagtagtgc cacacagcat aaaattagct tggtttcatg ctccgttaag tcatagcgac
     5581 taatcgctag ttcatttgct ttgaaaacaa ctaattcaga catacatctc aattggtcta
     5641 ggtgatttta atcactatac caattgagat gggctagtca atgataatta ctagtccttt
     5701 tcctttgagt tgtgggtatc tgtaaattct gctagacctt tgctggaaaa cttgtaaatt
     5761 ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt ttttttt
//
