LOCUS       sequence-43027-e        4795 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 4795)
  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 4795)
  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: pBbB7k-GFP
FEATURES             Location/Qualifiers
     source          1..4795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(8..1090)
                     /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     1052..1071
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1091..1168)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1374..1393
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1481..1500
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1481..1499
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1504..1520
                     /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             1561..2277
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1591..1619)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2214..2234
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2310..2381
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2397..2424
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2491..3153)
                     /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(3740..3834)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3865..4659)
                     /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(3976..3995)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4586..4605
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
       61 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
      121 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
      181 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
      241 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa
      301 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
      361 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
      421 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
      481 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
      541 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
      601 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
      661 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
      721 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
      781 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
      841 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
      901 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
      961 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     1021 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     1081 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     1141 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     1201 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     1261 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     1321 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     1381 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     1441 ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact cactataggg
     1501 gaattgtgag cggataacaa tttcagaatt caaaagatct tttaagaagg agatatacat
     1561 atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
     1621 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
     1681 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccgtg gccaacactt
     1741 gtcactactt tctcttatgg tgttcaatgc ttttcccgtt atccggatca catgaaacgg
     1801 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc
     1861 aaagatgacg ggaactacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt
     1921 aatcgtatcg agttaaaagg tattgatttt aaagaagatg gaaacattct cggacacaaa
     1981 ctggagtaca actataactc acacaatgta tacatcacgg cagacaaaca aaagaatgga
     2041 atcaaagcta acttcaaaat tcgccacaac attgaagatg gctccgttca actagcagac
     2101 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     2161 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt
     2221 cttgagtttg taactgctgc tgggattaca catggcatgg atgagctcta caaataagga
     2281 tccaaactcg agtaaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact
     2341 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     2401 caccttcggg tgggcctttc tgcgtttata cctaggctac agccgatagt ctggaacagc
     2461 gcacttacgg gttgctgcgc aacccaagtg ctaccggcgc ggcagcgtga cccgtgtcgg
     2521 cggctccaac ggctcgccat cgtccagaaa acacggctca tcgggcatcg gcaggcgctg
     2581 ctgcccgcgc cgttcccatt cctccgtttc ggtcaaggct ggcaggtctg gttccatgcc
     2641 cggaatgccg ggctggctgg gcggctcctc gccggggccg gtcggtagtt gctgctcgcc
     2701 cggatacagg gtcgggatgc ggcgcaggtc gccatgcccc aacagcgatt cgtcctggtc
     2761 gtcgtgatca accaccacgg cggcactgaa caccgacagg cgcaactggt cgcggggctg
     2821 gccccacgcc acgcggtcat tgaccacgta ggccaacacg gtgccggggc cgttgagctt
     2881 cacgacggag atccagcgct cggccaccaa gtccttgact gcgtattgga ccgtccgcaa
     2941 agaacgtccg atgagcttgg aaagtgtctt ctggctgacc accacggcgt tctggtggcc
     3001 catctgcgcc acgaggtgat gcagcagcat tgccgccgtg ggtttcctcg caataagccc
     3061 ggcccacgcc tcatgcgctt tgcgttccgt ttgcacccag tgaccgggct tgttcttggc
     3121 ttgaatgccg atttctctgg actgcgtggc catgcttatc tccatgcggt aggggtgccg
     3181 cacggttgcg gcaccatgcg caatcagctg caacttttcg gcagcgcgac aacaattatg
     3241 cgttgcgtaa aagtggcagt caattacaga ttttctttaa cctacgcaat gagctattgc
     3301 ggggggtgcc gcaatgagct gttgcgtacc cccctttttt aagttgttga tttttaagtc
     3361 tttcgcattt cgccctatat ctagttcttt ggtgcccaaa gaagggcacc cctgcggggt
     3421 tcccccacgc cttcggcgcg gctccccctc cggcaaaaag tggcccctcc ggggcttgtt
     3481 gatcgactgc gcggccttcg gccttgccca aggtggcgct gcccccttgg aacccccgca
     3541 ctcgccgccg tgaggctcgg ggggcaggcg ggcgggcttc gcccttcgac tgcccccact
     3601 cgcataggct tgggtcgttc caggcgcgtc aaggccaagc cgctgcgcgg tcgctgcgcg
     3661 agccttgacc cgccttccac ttggtgtcca accggcaagc gaagcgcgca ggccgcaggc
     3721 cggaggcact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     3781 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     3841 agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     3901 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     3961 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     4021 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     4081 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
     4141 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     4201 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     4261 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     4321 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     4381 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     4441 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     4501 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     4561 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     4621 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     4681 ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     4741 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attcc
//
