LOCUS       sequence-247748-        4209 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 4209)
  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 4209)
  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: pBbS12k-RFP
FEATURES             Location/Qualifiers
     source          1..4209
                     /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..764
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      797..868
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      884..911
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1442..2392)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(3154..3248)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3279..4073)
                     /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(3390..3409)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4000..4019
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcttga tggctagctc agtcctaggt acaatgctag caggtttaat cgaattcaaa
       61 agatctttta agaaggagat atacatatgg cgagtagcga agacgttatc aaagagttca
      121 tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg
      181 aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt accaaaggtg
      241 gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt tccaaagctt
      301 acgttaaaca cccggctgac atcccggact acctgaaact gtccttcccg gaaggtttca
      361 aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt taccgttacc caggactcct
      421 ccctgcaaga cggtgagttc atctacaaag ttaaactgcg tggtaccaac ttcccgtccg
      481 acggtccggt tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc
      541 cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc
      601 actacgacgc tgaagttaaa accacctaca tggctaaaaa accggttcag ctgccgggtg
      661 cttacaaaac cgacatcaaa ctggacatca cctcccacaa cgaagactac accatcgttg
      721 aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc ttaaggatcc aaactcgagt
      781 aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
      841 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
      901 gcctttctgc gtttatacct agggtacggg ttttgctgcc cgcaaacggg ctgttctggt
      961 gttgctagtt tgttatcaga atcgcagatc cggcttcagc cggtttgccg gctgaaagcg
     1021 ctatttcttc cagaattgcc atgatttttt ccccacggga ggcgtcactg gctcccgtgt
     1081 tgtcggcagc tttgattcga taagcagcat cgcctgtttc aggctgtcta tgtgtgactg
     1141 ttgagctgta acaagttgtc tcaggtgttc aatttcatgt tctagttgct ttgttttact
     1201 ggtttcacct gttctattag gtgttacatg ctgttcatct gttacattgt cgatctgttc
     1261 atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt
     1321 tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt
     1381 tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac
     1441 ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga
     1501 gccatgagaa cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct
     1561 caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt
     1621 tatgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct
     1681 ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca
     1741 acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta
     1801 aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat
     1861 tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag
     1921 ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt
     1981 caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag
     2041 gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg
     2101 tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca
     2161 tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca
     2221 tctgagcgta ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa
     2281 tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt
     2341 catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca
     2401 attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac
     2461 tagtcctttt cccgggtgat ctgggtatct gtaaattctg ctagaccttt gctggaaaac
     2521 ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt
     2581 atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag
     2641 atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt
     2701 cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca
     2761 ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg
     2821 agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg
     2881 ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa
     2941 gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc
     3001 tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt
     3061 atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac
     3121 aggcttaccc gtcttactgt ccctagtgct tggattctca ccaataaaaa acgcccggcg
     3181 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     3241 caggagtcca agcgagctct cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     3301 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     3361 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     3421 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     3481 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     3541 atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     3601 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     3661 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     3721 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     3781 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     3841 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctgcagttca
     3901 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     3961 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     4021 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
     4081 gatcctcatc ctgtctcttg atcagatcat gatcccctgc gccatcagat ccttggcggc
     4141 aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct
     4201 ggcaattcc
//
