LOCUS       sequence-230633-        5883 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 5883)
  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 5883)
  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..5883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(128..214)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     367..384
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(929..1720,1721..1789))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1552..1571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1790..1894)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1962..1980
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4271..4276,4277..4292,4293..4299)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     RBS             4323..4345
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(4353..4355,4356..5069)
                     /codon_start=1
                     /product="enhanced blue variant of GFP"
                     /label=EBFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKV
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     5003..5024
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5151..5171
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     rep_origin      complement(5302..5847)
                     /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(5382..5401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatctaaaac actaggcccc gtcaggatgg ccttctgctt aatttgatgc ctggcagttt
       61 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc aaatccgctc
      121 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
      181 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca
      241 tggggagacc ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgggg
      301 tcaggtggga ccaccgcgct actgccgcca ggcaaattct gttttatcag accgcttctg
      361 cgttctgatt taatctgtat caggctgaaa atcttctctc atccgccaaa acagccaagc
      421 tggatctggc aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct
      481 gtctttttcg tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat
      541 ggcactacag gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc
      601 ccgtcacggg cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt
      661 ttgctgttca gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt
      721 gacaggtcat tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta
      781 cccgtcttac tgtcaaccgg atctaaaaca ctagttaagg gattttggtc atgagattat
      841 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      901 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
      961 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1021 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1081 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1141 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1201 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1261 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1321 catggtcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1381 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1441 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1501 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     1561 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1621 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1681 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1741 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1801 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     1861 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     1921 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     1981 cctttcgtct tctagacaac atcagcaagg agaaaggggc taccggcgaa ccagcagccc
     2041 ctttataaag gcgcttcagt agtcagacca gcatcagtcc tgaaaaggcg ggcctgcgcc
     2101 cgcctccagg ttgctactta ccggattcgt aagccatgaa agccgccacc tccctgtgtc
     2161 cgtctctgta acgaatctcg cacagcgatt ttcgtgtcag ataagtgaat atcaacagtg
     2221 tgagacacac gatcaacaca caccagacaa gggaacttcg tggtagtttc atggccttct
     2281 tctccttgcg caaagcgcgg taagaggcta tcctgatgtg gactagacat agggatgcct
     2341 cgtggtggtt aatgaaaatt aacttactac ggggctatct tctttctgcc acacaacacg
     2401 gcaacaaacc accttcacgt catgaggcag aaagcctcaa gcgccgggca catcatagcc
     2461 catatacctg cacgctgacc acactcactt tccctgaaaa taatccgctc attcagaccg
     2521 ttcacgggaa atccgtgtga ttgttgccgc atcacgctgc ctcccggagt ttgtctcgag
     2581 cacttttgtt acccgccaaa caaaacccaa aaacaactca tacccaaccc aataaaacac
     2641 caaaacaaga caaataatca ttgattgatg gttgaaatgg ggtaaacttg acaaacaaac
     2701 ccacttaaaa cccaaaacat acccaaacac acaccaaaaa aacaccataa ggagttttat
     2761 aaatgttggt attcattgat gacggttcaa caaacatcaa actacagtag caggaaagcg
     2821 acggaacaat taaacagcac attagcccga acagcttcaa acgcgagtgg gcagtctctt
     2881 ttggtgataa aaaggtcttt aactacacac tgaacggcga acagtattca tttgatccaa
     2941 tcagcccgga tgctgtagtc acaaccaata tcgcatggca atacagcgac gttaatgtcg
     3001 ttgcagtgca tcacgcctta ctgaccagtg gtctgccggt aagcgaagtg gatattgttt
     3061 gcacacttcc tctgacagag tattacgaca gaaataacca acccaatacg gaaaatattg
     3121 agcgtaagaa agcaaacttc cggaaaaaaa ttacattaaa tggcggggat acattcacaa
     3181 taaaagatgt aaaagtcatg cctgaatcta taccggcagg ttatgaagtt ctacaagaac
     3241 tggatgagtt agattcttta ttaattatag atctcggggg caccacatta gatatttctc
     3301 aggtaatggg gaaattatcg gggatcagta aaatatacgg agactcatct cttggtgtct
     3361 ctctggttac atctgcagta aaagatgccc tttctcttgc gagaacaaaa ggaagtagct
     3421 atcttgctga cgatataatc attcacagaa aagataataa ctatctgaag caacgaatta
     3481 atgatgagaa cgaaatatca atagtcaccg aagcaatgaa tgaagcactt cgtaaacttg
     3541 agcaacgtgt attaaatacg ctcaatgaat tttctggtta tactcatgtt atggttatag
     3601 gcggtggcgc agaattaata tgcgatgcag taaaaaaaca cacacagatt cgtgatgaac
     3661 gttttttcaa aaccaataac tctcaatatg atttagttaa cggtatgtat ctcataggta
     3721 attaatgatg gacaagcgca gaaccattgc cttcaaacta aatccagatg taaatcaaac
     3781 agataaaatt gtttgtgata cactggacag tatcccgcaa ggggaacgaa gccgccttaa
     3841 ccgggccgca ctgacggcag gtctggcctt atacagacaa gatccccgga cccctttcct
     3901 tttatgtgag ctgctgacga aagaaaccac attttcagat atcgtgaata tattgagatc
     3961 gctatttcca aaagagatgg ccgattttaa ttcttcaata gtcactcaat cctcttcaca
     4021 acaagagcaa aaaagtgatg aagagaccaa aaaaaatgcg atgaagctaa taaattaatt
     4081 caattattat tgagttccct ttatccacta tcaggctgga taaagggaac tcaatcaagt
     4141 tattttctta ccagtcatta cataatcgtt attatgaaat aatcgtttgc actgtctctg
     4201 ttattcaggc aatttcaata aaggcacttg ctcacgctct gtcattttct gaaactctca
     4261 tgtggaattc ttgacaatta atcatcggct cgtataatgt gtggaggatc cctagaaata
     4321 attttgttta actttaagaa ggagatatac atatgagcaa gggcgaggag ctgttcaccg
     4381 gggtggtgcc catcctggtc gagctggatg gcgacgtaaa cggccacaag ttcagcgtgc
     4441 gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca
     4501 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     4561 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4621 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcacctac aagacccgcg
     4681 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4741 tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac agccacaacg
     4801 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4861 acgtcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4921 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgtc ctgagcaaag
     4981 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     5041 ctctcggcat ggacgagctg tacaagtgag cggccgcggg ccccatggat cgatagctgg
     5101 tcgacacaat ctgccctttc gaaagatccc aacgaaaagc gtgaccacat ggtccttctt
     5161 gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa ataatgagct
     5221 agcccgccta atgagcgggc ttttttttct cggcctagga gatacttaac agggaagtga
     5281 gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat
     5341 ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5401 ccctggcggc tcccccgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     5461 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     5521 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     5581 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     5641 ctggtaattg atttagagga gttagtcttg aagccatgcg ccggttaagg ctaaactgaa
     5701 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     5761 gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
     5821 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc
     5881 tag
//
