LOCUS       sequence-151988-        3070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promoter1 with BCD20-gfp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3070)
  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 3070)
  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: pProm1_BCD20-GFP
FEATURES             Location/Qualifiers
     source          1..3070
                     /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(1468..1470,1471..2184)
                     /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             1468..2181
                     /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(1498..1526)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2121..2141
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2217..2288
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2304..2331
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2374..2391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2508..3053)
                     /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(2588..2607)
                     /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 ataacaaaaa gagtattgac ttaaagtcta acctataggt ataatgtgtg
     1381 gagggcccaa gttcacttaa aaaggagatc aacaatgaaa gcaattttcg tactgaaaca
     1441 tcttaatcat gctgaggaaa gtttctaatg agcaaaggag aagaactttt cactggagtt
     1501 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtccgtgga
     1561 gagggtgaag gtgatgctac aaacggaaaa ctcaccctta aatttatttg cactactgga
     1621 aaactacctg ttccgtggcc aacacttgtc actactctga cctatggtgt tcaatgcttt
     1681 tcccgttatc cggatcacat gaaacggcat gactttttca agagtgccat gcccgaaggt
     1741 tatgtacagg aacgcactat atctttcaaa gatgacggga cctacaagac gcgtgctgaa
     1801 gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaagggtat tgattttaaa
     1861 gaagatggaa acattcttgg acacaaactc gagtacaact ttaactcaca caatgtatac
     1921 atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg ccacaacgtt
     1981 gaagatggtt ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc
     2041 cctgtccttt taccagacaa ccattacctg tcgacacaat ctgtcctttc gaaagatccc
     2101 aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat
     2161 ggcatggatg agctctacaa ataaggatcg gttgtcgagt aaggatctcc aggcatcaaa
     2221 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     2281 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatagga
     2341 tcctaactcg agcctaggga tatattccgc ttcctcgctc actgactcgc tacgctcggt
     2401 cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc
     2461 caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc
     2521 cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca
     2581 ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct
     2641 gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct
     2701 gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt
     2761 tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca
     2821 tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt
     2881 catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc
     2941 agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg
     3001 cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc
     3061 atcttattaa
//
