LOCUS       sequence-247095-        3321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression vector for Escherichia coli. BglBrick 
            compatible..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3321)
  AUTHORS   Jervis AJ, Carbonell P, Taylor S, Sung R, Dunstan MS, Robinson CJ, 
            Breitling R, Takano E, Scrutton NS
  TITLE     SelProm: A Queryable and Predictive Expression Vector Selection Tool
            for Escherichia coli.
  JOURNAL   ACS Synth Biol. 2019 Mar 18. doi: 10.1021/acssynbio.8b00399.
  PUBMED    30870592
REFERENCE   2  (bases 1 to 3321)
  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: "ACS Synth 
            Biol. 2019 Mar 18. doi: 10.1021/acssynbio.8b00399."
COMMENT     Sequence Label: pBbB20k-GFP
FEATURES             Location/Qualifiers
     source          1..3321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..41
                     /label=BBa_J23114
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.1"
     CDS             87..803
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(117..145)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     740..760
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      836..907
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      923..950
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1017..1679)
                     /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(2266..2360)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2391..3185)
                     /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(2502..2521)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3112..3131
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcttta tagctagctc agtcctaggt acaatgctag caggtttaat cgaattcaaa
       61 agatctttta agaaggagat atacatatga gtaaaggaga agaacttttc actggagttg
      121 tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag
      181 agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa
      241 aactacctgt tccgtggcca acacttgtca ctactttctc ttatggtgtt caatgctttt
      301 cccgttatcc ggatcacatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt
      361 atgtacagga acgcactata tctttcaaag atgacgggaa ctacaagacg cgtgctgaag
      421 tcaagtttga aggtgatacc cttgttaatc gtatcgagtt aaaaggtatt gattttaaag
      481 aagatggaaa cattctcgga cacaaactgg agtacaacta taactcacac aatgtataca
      541 tcacggcaga caaacaaaag aatggaatca aagctaactt caaaattcgc cacaacattg
      601 aagatggctc cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc
      661 ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca
      721 acgaaaagcg tgaccacatg gtccttcttg agtttgtaac tgctgctggg attacacatg
      781 gcatggatga gctctacaaa taaggatcca aactcgagta aggatctcca ggcatcaaat
      841 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
      901 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataccta
      961 ggctacagcc gatagtctgg aacagcgcac ttacgggttg ctgcgcaacc caagtgctac
     1021 cggcgcggca gcgtgacccg tgtcggcggc tccaacggct cgccatcgtc cagaaaacac
     1081 ggctcatcgg gcatcggcag gcgctgctgc ccgcgccgtt cccattcctc cgtttcggtc
     1141 aaggctggca ggtctggttc catgcccgga atgccgggct ggctgggcgg ctcctcgccg
     1201 gggccggtcg gtagttgctg ctcgcccgga tacagggtcg ggatgcggcg caggtcgcca
     1261 tgccccaaca gcgattcgtc ctggtcgtcg tgatcaacca ccacggcggc actgaacacc
     1321 gacaggcgca actggtcgcg gggctggccc cacgccacgc ggtcattgac cacgtaggcc
     1381 aacacggtgc cggggccgtt gagcttcacg acggagatcc agcgctcggc caccaagtcc
     1441 ttgactgcgt attggaccgt ccgcaaagaa cgtccgatga gcttggaaag tgtcttctgg
     1501 ctgaccacca cggcgttctg gtggcccatc tgcgccacga ggtgatgcag cagcattgcc
     1561 gccgtgggtt tcctcgcaat aagcccggcc cacgcctcat gcgctttgcg ttccgtttgc
     1621 acccagtgac cgggcttgtt cttggcttga atgccgattt ctctggactg cgtggccatg
     1681 cttatctcca tgcggtaggg gtgccgcacg gttgcggcac catgcgcaat cagctgcaac
     1741 ttttcggcag cgcgacaaca attatgcgtt gcgtaaaagt ggcagtcaat tacagatttt
     1801 ctttaaccta cgcaatgagc tattgcgggg ggtgccgcaa tgagctgttg cgtacccccc
     1861 ttttttaagt tgttgatttt taagtctttc gcatttcgcc ctatatctag ttctttggtg
     1921 cccaaagaag ggcacccctg cggggttccc ccacgccttc ggcgcggctc cccctccggc
     1981 aaaaagtggc ccctccgggg cttgttgatc gactgcgcgg ccttcggcct tgcccaaggt
     2041 ggcgctgccc ccttggaacc cccgcactcg ccgccgtgag gctcgggggg caggcgggcg
     2101 ggcttcgccc ttcgactgcc cccactcgca taggcttggg tcgttccagg cgcgtcaagg
     2161 ccaagccgct gcgcggtcgc tgcgcgagcc ttgacccgcc ttccacttgg tgtccaaccg
     2221 gcaagcgaag cgcgcaggcc gcaggccgga ggcactagtg cttggattct caccaataaa
     2281 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     2341 tggatctatc aacaggagtc caagcgagct ctcgaacccc agagtcccgc tcagaagaac
     2401 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     2461 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     2521 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     2581 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
     2641 tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     2701 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     2761 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     2821 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     2881 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     2941 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     3001 tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     3061 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     3121 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     3181 atcatgcgaa acgatcctca tcctgtctct tgatcagatc atgatcccct gcgccatcag
     3241 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     3301 ggcgccccag ctggcaattc c
//
