LOCUS       sequence-151993-        3059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promoter11 with BCD1-gfp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3059)
  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 3059)
  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: pProm11_BCD1-GFP
FEATURES             Location/Qualifiers
     source          1..3059
                     /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(1457..1459,1460..2173)
                     /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             1457..2170
                     /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(1487..1515)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2110..2130
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2206..2277
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2293..2320
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2363..2380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2497..3042)
                     /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(2577..2596)
                     /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 ataacttgac aattaatcat ccggctctta gtgtttgtgg agggcccaag
     1381 ttcacttaaa aaggagatca acaatgaaag caattttcgt actgaaacat cttaatcatg
     1441 cacaggagac tttctaatga gcaaaggaga agaacttttc actggagttg tcccaattct
     1501 tgttgaatta gatggtgatg ttaatgggca caaattttct gtccgtggag agggtgaagg
     1561 tgatgctaca aacggaaaac tcacccttaa atttatttgc actactggaa aactacctgt
     1621 tccgtggcca acacttgtca ctactctgac ctatggtgtt caatgctttt cccgttatcc
     1681 ggatcacatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     1741 acgcactata tctttcaaag atgacgggac ctacaagacg cgtgctgaag tcaagtttga
     1801 aggtgatacc cttgttaatc gtatcgagtt aaagggtatt gattttaaag aagatggaaa
     1861 cattcttgga cacaaactcg agtacaactt taactcacac aatgtataca tcacggcaga
     1921 caaacaaaag aatggaatca aagctaactt caaaattcgc cacaacgttg aagatggttc
     1981 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
     2041 accagacaac cattacctgt cgacacaatc tgtcctttcg aaagatccca acgaaaagcg
     2101 tgaccacatg gtccttcttg agtttgtaac tgctgctggg attacacatg gcatggatga
     2161 gctctacaaa taaggatcgg ttgtcgagta aggatctcca ggcatcaaat aaaacgaaag
     2221 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
     2281 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataggat cctaactcga
     2341 gcctagggat atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
     2401 ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac
     2461 ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga
     2521 caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag
     2581 ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt
     2641 taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt
     2701 ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc
     2761 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac
     2821 cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt
     2881 taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg
     2941 ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg
     3001 ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaa
//
