LOCUS       sequence-43018-e        5091 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 5091)
  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 5091)
  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: pBbB4k-GFP
FEATURES             Location/Qualifiers
     source          1..5091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1857..2573
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1887..1915)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2510..2530
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2606..2677
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2693..2720
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2787..3449)
                     /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(4036..4130)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4161..4955)
                     /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(4272..4291)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4882..4901
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtctcag cttttcagcc gccgccagaa cgtcgtccgg ctgatgccta aataattcgc
       61 cgctgctgtt ttatcgccat taaatttctc cagtgcctgt tgtggtgtca gtaagcgtgg
      121 agcgggagtt ttcgccgact cgcgcgccag ttccggcagt agcagctgca aaaattgcgg
      181 cgttaaatcc ggcgtcggtt ccacacttaa aaatagcgcc agtcgctcca tcatattgcg
      241 cagttcacga atattgcccg gccagtcgta gtgcaccagc acggtttcgc ttgcctgtaa
      301 tccctggcgt aatgcggcag aaaacggggt ggagagcgcc gccagagaca ctttcaaaaa
      361 gctttccgcc agtggcagaa tatccgccac ccgctcgcgc agtggtggca attgcagacg
      421 caaaatactc agccgataaa acaggtcacg gcgaaactgc ccttgccgca tatcttcttc
      481 cagattgcag tgagtggcgc taatgacccg cacatccacc ggaacaggct gatgcccgcc
      541 gacgcgggtg acctcttttt cttccagcac ccgcagcagc cgggtctgca acggcagcgg
      601 catttcgcca atctcatcga gaaacagcgt acctccgtgg gcgatttcaa acagcccggc
      661 gcgaccgccg cgtcgcgagc cggtaaacgc cccttcctca tagccaaaca gttctgcttc
      721 cagcagcgat tcggcaatcg ccccgcagtt gactgcaaca aacggatgcg actttttgcc
      781 ctgtcgcgca tcgtggcggg caaaatattc ccgatgaatc gcctgggccg ccagctcttt
      841 gcccgtcccc gtttccccct caatcaacac cgctgcactg gagcgggcat acagcaaaat
      901 agtctgccgt acttgttcca tctgtggtga ttgaccgagc atatcgccca gcacgtaacg
      961 agttctcagg gcgttgcggg tggcatcgtg agtgttatgg cgtaacgaca tgcgcgtcat
     1021 atccagcgca tcgctgaacg cctggcgcac ggtggcggcg gaatagataa aaattccggt
     1081 cattccggct tcttctgcca gatcggtaat cagccccgcg ccgaccaccg cttcggtgcc
     1141 gttagctttt agctcgttaa tctgcccgcg tgcgtcttct tcggtaatgt agctacgttg
     1201 atcgaggcgc aaattaaagg ttttttgaaa cgccaccagt gccggaatag tttcctgata
     1261 agtgacaacg ccgatagaag aggtgagttt tccggctttt gccagtgcct gtaacacatc
     1321 gtagccgctc ggtttaatca aaataactgg cactgacagg cggcttttca ggtacgcgcc
     1381 gttagatcca gcggcgatga tggcgtcaca gcgttcgttt gccagtttct tgtggatgta
     1441 ggtcaccgct ttttcaaagc caagctggat aggggtaatg ttcgccaggt gatcaaactc
     1501 aaggctgata tcgcgaaaca gctcgaacag gcgcgttaca gataccgtcc agataaccgg
     1561 tttgtcgtca ttaagccgtg gtggatgtgc catagcgcac cgcaaagtta agaaaccgaa
     1621 tattgggttt agtcttgttt cataattgtt gcaatgaaac gcggtgaaac attgcctgaa
     1681 acgttaactg aaacgcatat ttgcggatta gttcatgact ttatctctaa caaattgaaa
     1741 ttaaacattt aattttatta aggcaattgt ggcacacccc ttgctttgtc tttatcaacg
     1801 caaataacaa gttgataaca agaattcaaa agatctttta agaaggagat atacatatga
     1861 gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg
     1921 ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac
     1981 ttacccttaa atttatttgc actactggaa aactacctgt tccgtggcca acacttgtca
     2041 ctactttctc ttatggtgtt caatgctttt cccgttatcc ggatcacatg aaacggcatg
     2101 actttttcaa gagtgccatg cccgaaggtt atgtacagga acgcactata tctttcaaag
     2161 atgacgggaa ctacaagacg cgtgctgaag tcaagtttga aggtgatacc cttgttaatc
     2221 gtatcgagtt aaaaggtatt gattttaaag aagatggaaa cattctcgga cacaaactgg
     2281 agtacaacta taactcacac aatgtataca tcacggcaga caaacaaaag aatggaatca
     2341 aagctaactt caaaattcgc cacaacattg aagatggctc cgttcaacta gcagaccatt
     2401 atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt
     2461 ccacacaatc tgccctttcg aaagatccca acgaaaagcg tgaccacatg gtccttcttg
     2521 agtttgtaac tgctgctggg attacacatg gcatggatga gctctacaaa taaggatcca
     2581 aactcgagta aggatctcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     2641 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc
     2701 ttcgggtggg cctttctgcg tttataccta ggctacagcc gatagtctgg aacagcgcac
     2761 ttacgggttg ctgcgcaacc caagtgctac cggcgcggca gcgtgacccg tgtcggcggc
     2821 tccaacggct cgccatcgtc cagaaaacac ggctcatcgg gcatcggcag gcgctgctgc
     2881 ccgcgccgtt cccattcctc cgtttcggtc aaggctggca ggtctggttc catgcccgga
     2941 atgccgggct ggctgggcgg ctcctcgccg gggccggtcg gtagttgctg ctcgcccgga
     3001 tacagggtcg ggatgcggcg caggtcgcca tgccccaaca gcgattcgtc ctggtcgtcg
     3061 tgatcaacca ccacggcggc actgaacacc gacaggcgca actggtcgcg gggctggccc
     3121 cacgccacgc ggtcattgac cacgtaggcc aacacggtgc cggggccgtt gagcttcacg
     3181 acggagatcc agcgctcggc caccaagtcc ttgactgcgt attggaccgt ccgcaaagaa
     3241 cgtccgatga gcttggaaag tgtcttctgg ctgaccacca cggcgttctg gtggcccatc
     3301 tgcgccacga ggtgatgcag cagcattgcc gccgtgggtt tcctcgcaat aagcccggcc
     3361 cacgcctcat gcgctttgcg ttccgtttgc acccagtgac cgggcttgtt cttggcttga
     3421 atgccgattt ctctggactg cgtggccatg cttatctcca tgcggtaggg gtgccgcacg
     3481 gttgcggcac catgcgcaat cagctgcaac ttttcggcag cgcgacaaca attatgcgtt
     3541 gcgtaaaagt ggcagtcaat tacagatttt ctttaaccta cgcaatgagc tattgcgggg
     3601 ggtgccgcaa tgagctgttg cgtacccccc ttttttaagt tgttgatttt taagtctttc
     3661 gcatttcgcc ctatatctag ttctttggtg cccaaagaag ggcacccctg cggggttccc
     3721 ccacgccttc ggcgcggctc cccctccggc aaaaagtggc ccctccgggg cttgttgatc
     3781 gactgcgcgg ccttcggcct tgcccaaggt ggcgctgccc ccttggaacc cccgcactcg
     3841 ccgccgtgag gctcgggggg caggcgggcg ggcttcgccc ttcgactgcc cccactcgca
     3901 taggcttggg tcgttccagg cgcgtcaagg ccaagccgct gcgcggtcgc tgcgcgagcc
     3961 ttgacccgcc ttccacttgg tgtccaaccg gcaagcgaag cgcgcaggcc gcaggccgga
     4021 ggcactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa
     4081 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
     4141 ctcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     4201 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     4261 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
     4321 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     4381 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg
     4441 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     4501 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     4561 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     4621 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     4681 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     4741 gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg
     4801 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     4861 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     4921 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     4981 tgatcagatc atgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt
     5041 actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc c
//
