LOCUS       sequence-452514-        5548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Hybrid Driver Plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5548)
  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 5548)
  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: pZB573_pB1H1_T7-RNAP_LARP-I5CHO_1
FEATURES             Location/Qualifiers
     source          1..5548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(82..87,88..105,106..112)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    120..136
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     125..147
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     144..160
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     144..160
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(3348..4007)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3922..3941
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4008..4110)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4502..4519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4636..5181)
                     /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(4716..4735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5360..5378
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aaaaaaatat tgacaacata aaaaactttg tgttatactt gtaacgctac atggagatta
       61 actcaatcta gctagagagg ctttacactt tatgcttccg gctcgtataa tgtgtggaat
      121 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg
      181 aactctagac caaagagagg acacagtgtt gtttatcaag cctgcggatc tccgcgaaat
      241 tgtgactttt ccgctattta gcgatcttgt tcagtgtggc tttccttcac cggcagcaga
      301 ttacgttgaa cagcgcatcg atctgggtgg caacacgatt aacatcgcta agaacgactt
      361 ctctgacatc gaactggctg ctatcccgtt caacactctg gctgaccatt acggtgagcg
      421 tttagctcgc gaacagttgg cccttgagca tgagtcttac gagatgggtg aagcacgctt
      481 ccgcaagatg tttgagcgtc aacttaaagc tggtgaggtt gcggataacg ctgccgccaa
      541 gcctctcatc actaccctac tccctaagat gattgcacgc atcaacgact ggtttgagga
      601 agtgaaagct aagcgcggca agcgcccgac agccttccag ttcctgcaag aaatcaagcc
      661 ggaagccgta gcgtacatca ccattaagac cactctggct tgcctaacca gtgctgacaa
      721 tacaaccgtt caggctgtag caagcgcaat cggtcgggcc attgaggacg aggctcgctt
      781 cggtcgtatc cgtgaccttg aagctaagca cttcaagaaa aacgttgagg aacaactcaa
      841 caagcgcgta gggcacgtct acaagaaagc atttatgcaa gttgtcgagg ctgacatgct
      901 ctctaagggt ctactcggtg gcgaggcgtg gtcttcgtgg cataaggaag actctattca
      961 tgtaggagta cgctgcatcg agatgctcat tgagtcaacc ggaatggtta gcttacaccg
     1021 ccaaaatgct ggcgtagtag gtcaagactc tgagactatc gaactcgcac ctgaatacgc
     1081 tgaggctatc gcaacccgtg caggtgcgct ggctggcatc tctccgatgt tccaaccttg
     1141 cgtagttcct cctaagccgt ggactggcat tactggtggt ggctattggg ctaacggtcg
     1201 tcgtcctctg gcgctggtgc gtactcacag taagaaagca ctgatgcgct acgaagacgt
     1261 ttacatgcct gaggtgtaca aagcgattaa cattgcgcaa aacaccgcat ggaaaatcaa
     1321 caagaaagtc ctagcggtcg ccaacgtaat caccaagtgg aagcattgtc cggtcgagga
     1381 catccctgcg attgagcgtg aagaactccc gatgaaaccg gaagacatcg acatgaatcc
     1441 tgaggctctc accgcgtgga gacgtgctgc cgctgctgtg taccgcaagg acaaggctcg
     1501 caagtctcgc cgtatcagcc ttgagttcat gcttgagcaa gccaataagt ttgctaacca
     1561 taaggccatc tggttccctt acaacatgga ctggcgcggt cgtgtttacg ctgtgccaat
     1621 gttcaccccg caaggtaacg atatgaccaa aggactgctt acgctggcga aaggtaaacc
     1681 aatcggtaag gaaggttact actggctgaa aatccacggt gcaaactgtg cgggtgtcga
     1741 taaggttccg ttccctgagc gcatcaagtt cattgaggaa aaccacgaga acatcatggc
     1801 ttgcgctaag tctccactgg agaacacttg gtgggctgag caagattctc cgttctgctt
     1861 ccttgcgttc tgctttgagt acgctggggt acagcaccac ggcctgagct ataactgctc
     1921 ccttccgctg gcgtttgacg ggtcttgctc tggcatccag cacttctccg cgatgctccg
     1981 agatgaggta ggtggtcgcg cggttaactt gcttcctagt gaaaccgttc aggacatcta
     2041 cgggattgtt gctaagaaag tcaacgagat tctacaagca gacgcaatca atgggaccga
     2101 taacgaagta gttaccgtga ccgatgagaa cactggtgaa atctctgaga aagtcaagct
     2161 gggcactaag gcactggctg gtcaatggct ggcttacggt gttactcgca gtgtgactaa
     2221 gcgtccagtc atgacgctgg cttacgggtc caaagagttc ggcttccgtc aacaagtgct
     2281 ggaagatacc attcagccag ctattgattc cggcaagggt ctgatgttca ctcagccgaa
     2341 tcaggctgct ggatacatgg ctaagctgat ttgggaatct gtgagcgtga cggtggtagc
     2401 tgcggttgaa gcaatgaact ggcttaagtc tgctgctaag ctgctggctg ctgaggtcaa
     2461 agataagaag actggagaga ttcttcgcaa gcgttgcgct gtgcatggtg taactcctga
     2521 tggtttccct gtgtggtgcg aatacaagaa gcctattcag acgcgcttga acctgatgtt
     2581 cctcggtcag ttccgcttac agcctaccat taacaccaac aaagatagcg agattgatgc
     2641 acacaaacag gagtctggtc tggctcctaa ctttgtacac gcgcatgacg gtagccacct
     2701 tcgtaagact gtagtgtggg cacacgagaa gtacggaatc gaatcttttg cactgattca
     2761 cgactccttc ggtaccattc cggctgacgc tgcgaacctg ttcaaagcag tgcgcgaaac
     2821 tatggttgac acatatgagt cttgtgatgt actggctgat ttctacgacc agatcgctga
     2881 ccagttgcac gagtctcaat tggacaaaat gccagcactt ccggctaaag gtaacttgaa
     2941 cctccgtgac atcttagagt cggactacgc gttcgcgtaa taggatctgc atcgcaggat
     3001 gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctaa ccgtttttat
     3061 caggctctgg gaggcagaat aaatgatcat atcgtcaatt attacctcca cggggagagc
     3121 ctgagcaaac tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa
     3181 taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
     3241 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
     3301 gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
     3361 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     3421 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     3481 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     3541 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     3601 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     3661 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     3721 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     3781 gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     3841 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     3901 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     3961 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     4021 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     4081 tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag
     4141 ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc
     4201 acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt
     4261 atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc
     4321 agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag
     4381 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca
     4441 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
     4501 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     4561 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     4621 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
     4681 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4741 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     4801 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     4861 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     4921 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     4981 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     5041 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     5101 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     5161 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctagatt
     5221 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     5281 taattctcat gtttgacagc ttatcatcga taagctggac gtctaagaaa ccattattat
     5341 catgacatta acctataaaa ataggcgtat cacgaggccc tttggataac cagaagcaat
     5401 aaaaaatcaa atcggatttc actatataat ctcactttat ctaagatgaa tccgatggaa
     5461 gcatcctgtt ttctctcaat ttttttatct aaaacccagc gttcgatgct tctttgagcg
     5521 aacgatcaaa aataagtgcc ttcccatc
//
