LOCUS       sequence-151992-        3058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promoter9 with BCD1-gfp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3058)
  AUTHORS   Linshiz G, Stawski N, Goyal G, Bi C, Poust S, Sharma M, Mutalik V, 
            Keasling JD, Hillson NJ
  TITLE     PR-PR: cross-platform laboratory automation system.
  JOURNAL   ACS Synth Biol. 2014 Aug 15;3(8):515-24. doi: 10.1021/sb4001728. 
            Epub 2014 Jan 14.
  PUBMED    25126893
REFERENCE   2  (bases 1 to 3058)
  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/sb4001728";
            journalName: "ACS Synth Biol"; date: "2014-08-15- 15"; volume: "3"; 
            issue: "8"; pages: "515-24"
COMMENT     Sequence Label: pProm9_BCD1-GFP
FEATURES             Location/Qualifiers
     source          1..3058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(97..191)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(222..1016)
                     /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(333..352)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     943..962
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             join(1456..1458,1459..2172)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1456..2169
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1486..1514)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2109..2129
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2205..2276
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2292..2319
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2362..2379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2496..3041)
                     /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(2576..2595)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
       61 cagccccata cgatataagt tgttactagt gcttggattc tcaccaataa aaaacgcccg
      121 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
      181 caacaggagt ccaagcgagc tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
      241 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
      301 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
      361 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
      421 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
      481 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
      541 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
      601 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
      661 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
      721 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
      781 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
      841 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
      901 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
      961 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     1021 aacgatcctc atcctgtctc ttgatcagat catgatcccc tgcgccatca gatccttggc
     1081 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     1141 gctggcaatt ccgacgtcga attcaaaaga tcttaagtaa gtaagagtat acgtatatcg
     1201 gctaataacg tattaaggcg cttcggcgcc tttttttatg ggggtatttt catcccaatc
     1261 cacacgtcca acgcacagca aacaccacgt cgaccctatc agctgcgtgc tttctatgag
     1321 tcgttgctgc ataacttgcc tcttaatcat cggctcgtat aatgtgtgga gggcccaagt
     1381 tcacttaaaa aggagatcaa caatgaaagc aattttcgta ctgaaacatc ttaatcatgc
     1441 acaggagact ttctaatgag caaaggagaa gaacttttca ctggagttgt cccaattctt
     1501 gttgaattag atggtgatgt taatgggcac aaattttctg tccgtggaga gggtgaaggt
     1561 gatgctacaa acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt
     1621 ccgtggccaa cacttgtcac tactctgacc tatggtgttc aatgcttttc ccgttatccg
     1681 gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
     1741 cgcactatat ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt caagtttgaa
     1801 ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga agatggaaac
     1861 attcttggac acaaactcga gtacaacttt aactcacaca atgtatacat cacggcagac
     1921 aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacgttga agatggttcc
     1981 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
     2041 ccagacaacc attacctgtc gacacaatct gtcctttcga aagatcccaa cgaaaagcgt
     2101 gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag
     2161 ctctacaaat aaggatcggt tgtcgagtaa ggatctccag gcatcaaata aaacgaaagg
     2221 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     2281 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttataggatc ctaactcgag
     2341 cctagggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     2401 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
     2461 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
     2521 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
     2581 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
     2641 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
     2701 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
     2761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
     2821 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     2881 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     2941 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
     3001 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaa
//
