LOCUS       sequence-221570-        5915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGEN-mcs with a modular sfGFP expression scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5915)
  AUTHORS   Wiles TJ, Wall ES, Schlomann BH, Hay EA, Parthasarathy R, Guillemin 
            K
  TITLE     Modernized Tools for Streamlined Genetic Manipulation and 
            Comparative Study of Wild and Diverse Proteobacterial Lineages.
  JOURNAL   MBio. 2018 Oct 9;9(5). pii: mBio.01877-18. doi: 
            10.1128/mBio.01877-18.
  PUBMED    30301859
REFERENCE   2  (bases 1 to 5915)
  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: "MBio."; 
            date: "2018-10-9"; volume: "9(5). pii: mBio.01877-18. doi"; pages: "
            10.1128/mBio.01877-18"
COMMENT     Sequence Label: pXS-sfGFP
FEATURES             Location/Qualifiers
     source          1..5915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(39..66)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(158..244)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     397..414
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(959..1750,1751..1819))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1582..1601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1992..2010
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4303..4308,4309..4324,4325..4331)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     RBS             4355..4377
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(4385..4387,4388..5101)
                     /codon_start=1
                     /product="enhanced blue variant of GFP"
                     /label=EBFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKV
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     5035..5056
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5183..5203
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     rep_origin      complement(5334..5879)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(5414..5433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatctaaaac actaggccca agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
       61 ccttctgctt aatttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
      121 gccgttgctt cgcaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
      181 tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta
      241 tttgatgcct ggcagttccc tactctcgca tggggagacc ccacactacc atcggcgcta
      301 cggcgtttca cttctgagtt cggcatgggg tcaggtggga ccaccgcgct actgccgcca
      361 ggcaaattct gttttatcag accgcttctg cgttctgatt taatctgtat caggctgaaa
      421 atcttctctc atccgccaaa acagccaagc tggatctggc aaatcgctga atattccttt
      481 tgtctccgac catcaggcac ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc
      541 tcacggctct ggcagtgaat gggggtaaat ggcactacag gcgcctttta tggattcatg
      601 caaggaaact acccataata caagaaaagc ccgtcacggg cttctcaggg cgttttatgg
      661 cgggtctgct atgtggtgct atctgacttt ttgctgttca gcagttcctg ccctctgatt
      721 ttccagtctg accacttcgg attatcccgt gacaggtcat tcagactggc taatgcaccc
      781 agtaaggcag cggtatcatc aacaggctta cccgtcttac tgtcaaccgg atctaaaaca
      841 ctagttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1561 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1981 taacctataa aaataggcgt atcacgaggc cctttcgtct tctagacaac atcagcaagg
     2041 agaaaggggc taccggcgaa ccagcagccc ctttataaag gcgcttcagt agtcagacca
     2101 gcatcagtcc tgaaaaggcg ggcctgcgcc cgcctccagg ttgctactta ccggattcgt
     2161 aagccatgaa agccgccacc tccctgtgtc cgtctctgta acgaatctcg cacagcgatt
     2221 ttcgtgtcag ataagtgaat atcaacagtg tgagacacac gatcaacaca caccagacaa
     2281 gggaacttcg tggtagtttc atggccttct tctccttgcg caaagcgcgg taagaggcta
     2341 tcctgatgtg gactagacat agggatgcct cgtggtggtt aatgaaaatt aacttactac
     2401 ggggctatct tctttctgcc acacaacacg gcaacaaacc accttcacgt catgaggcag
     2461 aaagcctcaa gcgccgggca catcatagcc catatacctg cacgctgacc acactcactt
     2521 tccctgaaaa taatccgctc attcagaccg ttcacgggaa atccgtgtga ttgttgccgc
     2581 atcacgctgc ctcccggagt ttgtctcgag cacttttgtt acccgccaaa caaaacccaa
     2641 aaacaaccca tacccaaccc aataaaacac caaaacaaga caaataatca ttgattgatg
     2701 gttgaaatgg ggtaaacttg acaaacaaac ccacttaaaa cccaaaacat acccaaacac
     2761 acaccaaaaa aacaccataa ggagttttat aaatgttggt attcattgat gacggttcaa
     2821 caaacatcaa actacagtgg caggaaagcg acggaacaat taaacagcac attagcccga
     2881 acagcttcaa acgcgagtgg gcagtctctt ttggtgataa aaaggtcttt aactacacac
     2941 tgaacggcga acagtattca tttgatccaa tcagcccgga tgctgtagtc acaaccaata
     3001 tcgcatggca atacagcgac gttaatgtcg ttgcagtgca tcacgcctta ctgaccagtg
     3061 gtctgccggt aagcgaagtg gatattgttt gcacacttcc tctgacagag tattacgaca
     3121 gaaataacca acccaatacg gaaaatattg agcgtaagaa agcaaacttc cggaaaaaaa
     3181 ttacattaaa tggcggggat acattcacaa taaaagatgt aaaagtcatg cctgaatcta
     3241 taccggcagg ttatgaagtt ctacaagaac tggatgagtt agattcttta ttaattatag
     3301 atctcggggg caccacatta gatatttctc aggtaatggg gaaattatcg gggatcagta
     3361 aaatatacgg agactcatct cttggtgtct ctctggttac atctgcagta aaagatgccc
     3421 tttctcttgc gagaacaaaa ggaagtagct atcttgctga cgatataatc attcacagaa
     3481 aagataataa ctatctgaag caacgaatta atgatgagaa caaaatatca atagtcaccg
     3541 aagcaatgaa tgaagcactt cgtaaacttg agcaacgtgt attaaatacg ctcaatgaat
     3601 tttctggtta tactcatgtt atggttatag gcggtggcgc agaattaata tgcgatgcag
     3661 taaaaaaaca cacacagatt cgtgatgaac gttttttcaa aaccaataac tctcaatatg
     3721 atttagttaa cggtatgtat ctcataggta attaatgatg gacaagcgca gaaccattgc
     3781 cttcaaacta aatccagatg taaatcaaac agataaaatt gtttgtgata cactggacag
     3841 tatcccgcaa ggggaacgaa gccgccttaa ccgggccgca ctgacggcag gtctggcctt
     3901 atacagacaa gatccccgga cccctttcct tttatgtgag ctgctgacga aagaaaccac
     3961 attttcagat atcgtgaata tattgagatc gctatttcca aaagagatgg ccgattttaa
     4021 ttcttcaata gtcactcaat cctcttcaca acaagagcaa aaaagtgatg aagagaccaa
     4081 aaaaaatgcg atgaagctaa taaattaatt caattattat tgagttccct ttatccacta
     4141 tcaggctgga taaagggaac tcaatcaagt tattttctta ccagtcatta cataatcgtt
     4201 attatgaaat aatcgtttgc actgtctctg ttattcaggc aatttcaata aaggcacttg
     4261 ctcacgctct gtcattttct gaaactcttc atgctggaat tcttgacaat taatcatcgg
     4321 ctcgtataat gtgtggagga tccctagaaa taattttgtt taactttaag aaggagatat
     4381 acatatgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     4441 tggcgacgta aacggccaca agttcagcgt gcgcggcgag ggcgagggcg atgccaccaa
     4501 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     4561 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     4621 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     4681 cttcaaggac gacggcacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     4741 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     4801 caagctggag tacaacttca acagccacaa cgtctatatc atggccgaca agcagaagaa
     4861 cggcatcaag gtgaacttca agatccgcca caacgtcgag gacggcagcg tgcagctcgc
     4921 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     4981 ctacctgagc acccagtccg tcctgagcaa agaccccaac gagaagcgcg atcacatggt
     5041 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtg
     5101 agcggccgcg ggccccatgg atcgatagct ggtcgacaca atctgccctt tcgaaagatc
     5161 ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct gggattacac
     5221 atggcatgga tgagctctac aaataatgag ctagcccgcc taatgagcgg gctttttttt
     5281 ctcggcctag gagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat
     5341 aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac
     5401 ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct
     5461 gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc
     5521 acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc
     5581 ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga
     5641 aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct
     5701 tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc
     5761 caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct
     5821 gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag
     5881 aagatcatct tattaatcag ataaaatatt tctag
//
