LOCUS       sequence-117449-        5176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  OR2-OR1-P2 promoter expressing T7 RNAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5176)
  AUTHORS   Siegal-Gaskins D, Tuza ZA, Kim J, Noireaux V, Murray RM
  TITLE     Gene circuit performance characterization and resource usage in a 
            cell-free ""breadboard"".
  JOURNAL   ACS Synth Biol. 2014 Jun 20;3(6):416-25. doi: 10.1021/sb400203p. 
            Epub 2014 Apr 7.
  PUBMED    24670245
REFERENCE   2  (bases 1 to 5176)
  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/sb400203p";
            journalName: "ACS Synth Biol"; date: "2014-06-20- 20"; volume: "3"; 
            issue: "6"; pages: "416-25"
COMMENT     Sequence Label: Pr2-T7RNAP
FEATURES             Location/Qualifiers
     source          1..5176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..24
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             362..384
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3219..3236)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3390..3978)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3470..3489)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4149..4940,4941..5009))
                     /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     4772..4791
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5010..5114)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ataaaaatag gcgtatcacg aggccctttc gtcttcaaga attctggcga atcctctgac
       61 cagccagaaa acgacctttc tgtggtgaaa ccggatgctg caattcagag cggcagcaag
      121 tgggggacag cagaagacct gaccgccgca gagtggatgt ttgacatggt gaagactatc
      181 gcaccatcag ccagaaaacc gaattttgct gggtgggcta acgatatccg cctgatgcgt
      241 gaacgtgacg gacgtaacca ccgcgacatg tgtgtgctgt tccgctgggc atgctgagct
      301 aacaccgtgc gtgttgacaa ttttacctct ggcggtgcta atggttgcag ctagcaataa
      361 ttttgtttaa ctttaagaag gagatatacc atgaacacga ttaacatcgc taagaacgac
      421 ttctctgaca tcgaactggc tgctatcccg ttcaacactc tggctgacca ttacggtgag
      481 cgtttagctc gcgaacagtt ggcccttgag catgagtctt acgagatggg tgaagcacgc
      541 ttccgcaaga tgtttgagcg tcaacttaaa gctggtgagg ttgcggataa cgctgccgcc
      601 aagcctctca tcactaccct actccctaag atgattgcac gcatcaacga ctggtttgag
      661 gaagtgaaag ctaagcgcgg caagcgcccg acagccttcc agttcctgca agaaatcaag
      721 ccggaagccg tagcgtacat caccattaag accactctgg cttgcctaac cagtgctgac
      781 aatacaaccg ttcaggctgt agcaagcgca atcggtcggg ccattgagga cgaggctcgc
      841 ttcggtcgta tccgtgacct tgaagctaag cacttcaaga aaaacgttga ggaacaactc
      901 aacaagcgcg tagggcacgt ctacaagaaa gcatttatgc aagttgtcga ggctgacatg
      961 ctctctaagg gtctactcgg tggcgaggcg tggtcttcgt ggcataagga agactctatt
     1021 catgtaggag tacgctgcat cgagatgctc attgagtcaa ccggaatggt tagcttacac
     1081 cgccaaaatg ctggcgtagt aggtcaagac tctgagacta tcgaactcgc acctgaatac
     1141 gctgaggcta tcgcaacccg tgcaggtgcg ctggctggca tctctccgat gttccaacct
     1201 tgcgtagttc ctcctaagcc gtggactggc attactggtg gtggctattg ggctaacggt
     1261 cgtcgtcctc tggcgctggt gcgtactcac agtaagaaag cactgatgcg ctacgaagac
     1321 gtttacatgc ctgaggtgta caaagcgatt aacattgcgc aaaacaccgc atggaaaatc
     1381 aacaagaaag tcctagcggt cgccaacgta atcaccaagt ggaagcattg tccggtcgag
     1441 gacatccctg cgattgagcg tgaagaactc ccgatgaaac cggaagacat cgacatgaat
     1501 cctgaggctc tcaccgcgtg gaaacgtgct gccgctgctg tgtaccgcaa ggacaaggct
     1561 cgcaagtctc gccgtatcag ccttgagttc atgcttgagc aagccaataa gtttgctaac
     1621 cataaggcca tctggttccc ttacaacatg gactggcgcg gtcgtgttta cgctgtgtca
     1681 atgttcaacc cgcaaggtaa cgatatgacc aaaggactgc ttacgctggc gaaaggtaaa
     1741 ccaatcggta aggaaggtta ctactggctg aaaatccacg gtgcaaactg tgcgggtgtc
     1801 gataaggttc cgttccctga gcgcatcaag ttcattgagg aaaaccacga gaacatcatg
     1861 gcttgcgcta agtctccact ggagaacact tggtgggctg agcaagattc tccgttctgc
     1921 ttccttgcgt tctgctttga gtacgctggg gtacagcacc acggcctgag ctataactgc
     1981 tcccttccgc tggcgtttga cgggtcttgc tctggcatcc agcacttctc cgcgatgctc
     2041 cgagatgagg taggtggtcg cgcggttaac ttgcttccta gtgaaaccgt tcaggacatc
     2101 tacgggattg ttgctaagaa agtcaacgag attctacaag cagacgcaat caatgggacc
     2161 gataacgaag tagttaccgt gaccgatgag aacactggtg aaatctctga gaaagtcaag
     2221 ctgggcacta aggcactggc tggtcaatgg ctggcttacg gtgttactcg cagtgtgact
     2281 aagcgttcag tcatgacgct ggcttacggg tccaaagagt tcggcttccg tcaacaagtg
     2341 ctggaagata ccattcagcc agctattgat tccggcaagg gtctgatgtt cactcagccg
     2401 aatcaggctg ctggatacat ggctaagctg atttgggaat ctgtgagcgt gacggtggta
     2461 gctgcggttg aagcaatgaa ctggcttaag tctgctgcta agctgctggc tgctgaggtc
     2521 aaagataaga agactggaga gattcttcgc aagcgttgcg ctgtgcattg ggtaactcct
     2581 gatggtttcc ctgtgtggca ggaatacaag aagcctattc agacgcgctt gaacctgatg
     2641 ttcctcggtc agttccgctt acagcctacc attaacacca acaaagatag cgagattgat
     2701 gcacacaaac aggagtctgg tatcgctcct aactttgtac acagccaaga cggtagccac
     2761 cttcgtaaga ctgtagtgtg ggcacacgag aagtacggaa tcgaatcttt tgcactgatt
     2821 cacgactcct tcggtaccat tccggctgac gctgcgaacc tgttcaaagc agtgcgcgaa
     2881 actatggttg acacatatga gtcttgtgat gtactggctg atttctacga ccagttcgct
     2941 gaccagttgc acgagtctca attggacaaa atgccagcac ttccggctaa aggtaacttg
     3001 aacctccgtg acatcttaga gtcggacttc gcgttcgcgt aactcgagca aagcccgccg
     3061 aaaggcgggc ttttctgtgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     3121 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     3181 aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc
     3241 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3301 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3361 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3421 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3481 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3541 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     3601 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3661 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3721 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3781 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     3841 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3901 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3961 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4021 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4081 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4141 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4201 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4261 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4321 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4381 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4441 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4501 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4561 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4621 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4681 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4741 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4801 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4861 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4921 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4981 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5041 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5101 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5161 atcatgacat taacct
//
