LOCUS       sequence-43009-e        4727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4727)
  AUTHORS   Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, 
            Keasling JD
  TITLE     BglBrick vectors and datasheets: A synthetic biology platform for 
            gene expression.
  JOURNAL   J Biol Eng. 2011 Sep 20;5:12.
  PUBMED    21933410
REFERENCE   2  (bases 1 to 4727)
  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"; journalName: "J Biol 
            Eng."; date: "2011-09-20"; volume: "5"; pages: "12"
COMMENT     Sequence Label: pBbB1k-GFP
FEATURES             Location/Qualifiers
     source          1..4727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..84
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             85..1167
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(104..123)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1399..1404,1405..1421,1422..1428)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1436..1452
                     /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)."
     CDS             1493..2209
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1523..1551)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2146..2166
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2242..2313
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2329..2356
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2423..3085)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVLAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     terminator      complement(3672..3766)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3797..4591)
                     /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(3908..3927)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4518..4537
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
       61 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
      121 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
      181 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
      241 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
      301 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
      361 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
      421 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
      481 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
      541 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
      601 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
      661 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
      721 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
      781 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
      841 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
      901 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
      961 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     1021 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     1081 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1141 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agcgcgaatt
     1201 gatctggttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca
     1261 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     1321 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     1381 atattctgaa atgagctgtt gacaattaat catccggctc gtataatgtg tggaattgtg
     1441 agcggataac aatttcagaa ttcaaaagat cttttaagaa ggagatatac atatgagtaa
     1501 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     1561 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     1621 ccttaaattt atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac
     1681 tttctcttat ggtgttcaat gcttttcccg ttatccggat cacatgaaac ggcatgactt
     1741 tttcaagagt gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga
     1801 cgggaactac aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat
     1861 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt ctcggacaca aactggagta
     1921 caactataac tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc
     1981 taacttcaaa attcgccaca acattgaaga tggctccgtt caactagcag accattatca
     2041 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     2101 acaatctgcc ctttcgaaag atcccaacga aaagcgtgac cacatggtcc ttcttgagtt
     2161 tgtaactgct gctgggatta cacatggcat ggatgagctc tacaaataag gatccaaact
     2221 cgagtaagga tctccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     2281 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     2341 ggtgggcctt tctgcgttta tacctaggct acagccgata gtctggaaca gcgcacttac
     2401 gggttgctgc gcaacccaag tgctaccggc gcggcagcgt gacccgtgtc ggcggctcca
     2461 acggctcgcc atcgtccaga aaacacggct catcgggcat cggcaggcgc tgctgcccgc
     2521 gccgttccca ttcctccgtt tcggtcaagg ctggcaggtc tggttccatg cccggaatgc
     2581 cgggctggct gggcggctcc tcgccggggc cggtcggtag ttgctgctcg cccggataca
     2641 gggtcgggat gcggcgcagg tcgccatgcc ccaacagcga ttcgtcctgg tcgtcgtgat
     2701 caaccaccac ggcggcactg aacaccgaca ggcgcaactg gtcgcggggc tggccccacg
     2761 ccacgcggtc attgaccacg taggccaaca cggtgccggg gccgttgagc ttcacgacgg
     2821 agatccagcg ctcggccacc aagtccttga ctgcgtattg gaccgtccgc aaagaacgtc
     2881 cgatgagctt ggaaagtgtc ttctggctga ccaccacggc gttctggtgg cccatctgcg
     2941 ccacgaggtg atgcagcagc attgccgccg tgggtttcct cgcaataagc ccggcccacg
     3001 cctcatgcgc tttgcgttcc gtttgcaccc agtgaccggg cttgttcttg gcttgaatgc
     3061 cgatttctct ggactgcgtg gccatgctta tctccatgcg gtaggggtgc cgcacggttg
     3121 cggcaccatg cgcaatcagc tgcaactttt cggcagcgcg acaacaatta tgcgttgcgt
     3181 aaaagtggca gtcaattaca gattttcttt aacctacgca atgagctatt gcggggggtg
     3241 ccgcaatgag ctgttgcgta cccccctttt ttaagttgtt gatttttaag tctttcgcat
     3301 ttcgccctat atctagttct ttggtgccca aagaagggca cccctgcggg gttcccccac
     3361 gccttcggcg cggctccccc tccggcaaaa agtggcccct ccggggcttg ttgatcgact
     3421 gcgcggcctt cggccttgcc caaggtggcg ctgccccctt ggaacccccg cactcgccgc
     3481 cgtgaggctc ggggggcagg cgggcgggct tcgcccttcg actgccccca ctcgcatagg
     3541 cttgggtcgt tccaggcgcg tcaaggccaa gccgctgcgc ggtcgctgcg cgagccttga
     3601 cccgccttcc acttggtgtc caaccggcaa gcgaagcgcg caggccgcag gccggaggca
     3661 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     3721 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctctcg
     3781 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     3841 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     3901 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     3961 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     4021 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
     4081 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     4141 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     4201 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     4261 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     4321 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     4381 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     4441 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     4501 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     4561 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     4621 cagatcatga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
     4681 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattcc
//
