LOCUS       sequence-43015-e        5156 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 5156)
  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 5156)
  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: pBbB3k-GFP
FEATURES             Location/Qualifiers
     source          1..5156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1922..2638
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1952..1980)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2575..2595
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2671..2742
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2758..2785
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2852..3514)
                     /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(4101..4195)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4226..5020)
                     /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(4337..4356)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4947..4966
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcttaa ttacccgact ggtctttggc accagctttt aaacgacgcc acagagtggt
       61 gcggctaatc cccagataac gcgccgcggc ggtcttatcg cccttaaagc gcgccagtac
      121 atcctgtaac gcgttcgcat ccacggttga aggcgtcagt tctgctgtgt tcaccataag
      181 ctcaggtaat aactgccgca taaattgcct gtccagcgtt ggcgcgggat cgacgcttaa
      241 aaaaagcgcc aggcgctcca tcatattacg cagttcgcga atattaccgg gccagcgcca
      301 ggccagcaaa agcggctgac actgtgtcaa tccatgacgt atcgattcgg taaacggaat
      361 ttccatcgcc gccagcgact gttttaaaaa gctttccgcc agcggcaaaa tatcagcctg
      421 ccgctcgcgc aaaggaggaa gcgtcagacg cagaatactc aggcgataaa agagatcggg
      481 gcgaaaacgt ccttgcatta tctcccgatc cagatcgcaa tgcgtagcgc tgatcacccg
      541 gacatccacc gggatcggct ggtgtccgcc gacgcgagtg acggcttttt cctccagtac
      601 gcgcaaaagt cgggtttgta acggcaaggg catttcgccg atttcatcca gaaacagcgt
      661 gccgccgtgt gcgatttcaa acagccccgc ccggcctcct cgtcgtgaac cggtaaacgc
      721 gccctcttca taaccaaaca gttccgcttc cagcaacgac tcggtaatcg cgccgcaatt
      781 gacggcgaca aagggagggg atggcttatt ctgacggtga ggctggcggt gaaagaacgt
      841 ctggtgaatc gcctgcgccg ccagctcttt tccggtccct gtttccccct gaatcagcac
      901 tgccgcacgg gagcgggcat agagcgtaat cgtctggcgg agctgctcca tttgcggcga
      961 ctggccgcgt atatcgccca gttcataccg ggtttgtaat cccttgccgg atgggtaatc
     1021 cacgcgctgg cgccgtgtca gacgggtcat atccagcgca tcatgaaaag cctgacgaac
     1081 ggtcgccgcg gaataaataa agatggcggt cattcctgcc tcttccgcca gatcggtaat
     1141 taatcccgcg ccgacgacgg cctcaatacc gttggcctta agttcgttaa tttgcccgcg
     1201 cgcgtcctct tcggtgacat agcttcgctg ttcaagacgg aggtgaaacg ttttctgaaa
     1261 ggcaagtaaa gccggaatgg tctcctgata ggtcacgata ccgatagacg aggtgagctt
     1321 tcccgctttc gccagcgcct gtaatacatc gaatccgctg ggcttgatga ggatcaccgg
     1381 tattgacagg cggcttttta aataggcccc attcgaaccc gccgcgataa tcgcgtcgca
     1441 gcgctcggtc gccagttttt tgcgaatgta ggtcaccgcc ttttcaaagc cgagttgaat
     1501 aggcgtgatg gtcgccagat gatcaaattc caggctgata tcccgaaaca gttcgaacag
     1561 acgcgttacg gagaccgtcc agatcaccgg tttatcgcta ttatcgcgcg gagcgctgtg
     1621 ggcagtcgtc atcgcagtag attcatcttt aaggggcgat tttttgtttt aaacgtgttt
     1681 cataaatgtt gcaatgaaac agggtgattc gtttcatgaa acgttagctg acacgttttt
     1741 ttttccctta atcgcgctta ttcataacag aaatgactgt aattacctgt ttttaaatct
     1801 cattgtattt aattttctcg ctgcctcttt gcctggcata gcctttgctt tggtgaatac
     1861 atcttgaata acaatttact aacatggaat tcaaaagatc ttttaagaag gagatataca
     1921 tatgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
     1981 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
     2041 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccgt ggccaacact
     2101 tgtcactact ttctcttatg gtgttcaatg cttttcccgt tatccggatc acatgaaacg
     2161 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatcttt
     2221 caaagatgac gggaactaca agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt
     2281 taatcgtatc gagttaaaag gtattgattt taaagaagat ggaaacattc tcggacacaa
     2341 actggagtac aactataact cacacaatgt atacatcacg gcagacaaac aaaagaatgg
     2401 aatcaaagct aacttcaaaa ttcgccacaa cattgaagat ggctccgttc aactagcaga
     2461 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
     2521 cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagcgtgacc acatggtcct
     2581 tcttgagttt gtaactgctg ctgggattac acatggcatg gatgagctct acaaataagg
     2641 atccaaactc gagtaaggat ctccaggcat caaataaaac gaaaggctca gtcgaaagac
     2701 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
     2761 tcaccttcgg gtgggccttt ctgcgtttat acctaggcta cagccgatag tctggaacag
     2821 cgcacttacg ggttgctgcg caacccaagt gctaccggcg cggcagcgtg acccgtgtcg
     2881 gcggctccaa cggctcgcca tcgtccagaa aacacggctc atcgggcatc ggcaggcgct
     2941 gctgcccgcg ccgttcccat tcctccgttt cggtcaaggc tggcaggtct ggttccatgc
     3001 ccggaatgcc gggctggctg ggcggctcct cgccggggcc ggtcggtagt tgctgctcgc
     3061 ccggatacag ggtcgggatg cggcgcaggt cgccatgccc caacagcgat tcgtcctggt
     3121 cgtcgtgatc aaccaccacg gcggcactga acaccgacag gcgcaactgg tcgcggggct
     3181 ggccccacgc cacgcggtca ttgaccacgt aggccaacac ggtgccgggg ccgttgagct
     3241 tcacgacgga gatccagcgc tcggccacca agtccttgac tgcgtattgg accgtccgca
     3301 aagaacgtcc gatgagcttg gaaagtgtct tctggctgac caccacggcg ttctggtggc
     3361 ccatctgcgc cacgaggtga tgcagcagca ttgccgccgt gggtttcctc gcaataagcc
     3421 cggcccacgc ctcatgcgct ttgcgttccg tttgcaccca gtgaccgggc ttgttcttgg
     3481 cttgaatgcc gatttctctg gactgcgtgg ccatgcttat ctccatgcgg taggggtgcc
     3541 gcacggttgc ggcaccatgc gcaatcagct gcaacttttc ggcagcgcga caacaattat
     3601 gcgttgcgta aaagtggcag tcaattacag attttcttta acctacgcaa tgagctattg
     3661 cggggggtgc cgcaatgagc tgttgcgtac cccccttttt taagttgttg atttttaagt
     3721 ctttcgcatt tcgccctata tctagttctt tggtgcccaa agaagggcac ccctgcgggg
     3781 ttcccccacg ccttcggcgc ggctccccct ccggcaaaaa gtggcccctc cggggcttgt
     3841 tgatcgactg cgcggccttc ggccttgccc aaggtggcgc tgcccccttg gaacccccgc
     3901 actcgccgcc gtgaggctcg gggggcaggc gggcgggctt cgcccttcga ctgcccccac
     3961 tcgcataggc ttgggtcgtt ccaggcgcgt caaggccaag ccgctgcgcg gtcgctgcgc
     4021 gagccttgac ccgccttcca cttggtgtcc aaccggcaag cgaagcgcgc aggccgcagg
     4081 ccggaggcac tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt
     4141 ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc
     4201 gagctctcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
     4261 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     4321 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     4381 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     4441 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga
     4501 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     4561 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     4621 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     4681 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     4741 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     4801 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
     4861 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     4921 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     4981 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     5041 tctcttgatc agatcatgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     5101 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattcc
//
