LOCUS       sequence-240706-        4176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 promoter expressing deGFP-MG aptamer fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4176)
  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 4176)
  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: PT7-deGFP-MGapt
FEATURES             Location/Qualifiers
     source          1..4176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(110..131)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(371..390)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     718..739
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(860..878)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      874..921
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1287..1292,1293..1309,1310..1315))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1336..1352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1336..1352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1349..1371)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1360..1376
                     /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)."
     promoter        complement(join(1384..1390,1391..1408,1409..1414))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1429..1450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1567..1584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1738..2326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1818..1837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2497..3288,3289..3357))
                     /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     3120..3139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3358..3462)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3530..3548
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3586..3608)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3708..3727
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3923..3945
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3937..3954
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3938..3954
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4074..4093
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacca tggagctttt cactggcgtt gttcccatcc tggtcgagct
      121 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      181 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      241 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      301 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      361 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      421 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
      481 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
      541 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
      601 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
      661 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
      721 ggtcctgctg gagttcgtga ccgccgccgg gatctaactc gagccttagg agatccggct
      781 gctaacggga tcccgactgg cgagagccag gtaacgaatg gatccaaagc ccgaaaggaa
      841 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      901 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc acaggacggg
      961 tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga gcaggactgg
     1021 gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa attgcatcaa
     1081 cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat ggacgatatc
     1141 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac
     1201 ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta
     1261 gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg
     1321 agaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1381 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1441 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1501 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1561 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1621 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1681 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1741 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1801 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1861 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1921 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1981 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2041 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2101 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2161 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2221 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2281 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2341 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2401 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2461 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2521 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2581 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2641 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2701 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2761 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2821 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2881 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2941 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3001 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3061 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3121 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3181 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3241 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3301 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3361 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3421 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3481 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3541 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3601 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3661 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     3721 gattgtactg agagtgcacc atatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     3781 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     3841 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     3901 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag
     3961 cttgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     4021 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     4081 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     4141 ccggcgtaga ggatcgagat ctcgatcccg cgaaat
//
