LOCUS       pPM_Pveg-sfgfp                7666 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Integration vector that expresses gfp in S. mutans.
ACCESSION   .
VERSION     .
KEYWORDS    pPM_Pveg-sfgfp
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7666)
  AUTHORS   Shields RC, Kaspar JR, Lee K, Underhill SAM, Burne RA
  TITLE     Fluorescence tools adapted for real-time monitoring of the behaviors
            of Streptococcus species.
  JOURNAL   Appl Environ Microbiol. 2019 May 17. pii: AEM.00620-19. doi: 
            10.1128/AEM.00620-19.
  PUBMED    31101614
REFERENCE   2  (bases 1 to 7666)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..7666
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             596..1312
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(1766..2560)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     primer_bind     complement(4107..4123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4107..4123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4120..4142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4131..4147
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4155..4185)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4200..4221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4458..4475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4592..5137)
                     /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(4672..4691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7077..7096
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7163..7265)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     7652..8
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7666..17
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttctagaagg cgaatatttt taacacgtgt tccttgtttt
       61 aaatctttag gagccccctt aactttaaga tccttaataa gagtgactgt atcaccatca
      121 gctaattgtt tgccattcgc atcaataaca acaagtcctt cttctttctc aacttctgca
      181 ggattccatt cataagcaca ttctggacaa accaatagaa taccatcttc atagacatac
      241 tctgactgac atttaggaca attaggtaaa gacatataaa ctccttttat tttgattacc
      301 attaacagtt tacctgagtt tttatcagtt ggcaagtctt atcacaacaa aaagctagct
      361 gcttgtactg actctatgag ttaagtatac tgtctaactt atagaaaacg cagaacaaga
      421 gctagttctt ttaattataa gagtttccct taatctaaga ttcaacactg gatctccaac
      481 gcgttgggag ctctagatcg aattccttat taacgttgat ataatttaaa ttttatttga
      541 caaaaatggg ctcgtgttgt acaataaatg tgattaacta ataaggagga caaacatgtc
      601 aaaaggagaa gagctgttca caggtgttgt gccgattctc gttgagcttg acggagatgt
      661 aaacggacac aaattctctg ttcgcggtga aggtgaagga gatgcaacaa acggcaagct
      721 gacattgaag tttatttgca caactggaaa gctgccggtt ccttggccga cacttgtaac
      781 gacgctgact tacggcgttc aatgcttctc tcgttatcca gaccacatga aacgccatga
      841 tttcttcaaa tctgcaatgc ctgaaggcta cgttcaagag cgtacgatca gcttcaaaga
      901 tgacggaacg tacaaaacaa gagcagaagt gaagtttgaa ggtgacacac ttgtgaaccg
      961 cattgaattg aaaggcattg atttcaaaga agatggaaac atccttggac acaaacttga
     1021 atacaacttc aacagccaca acgtatacat cactgctgac aaacaaaaaa acggcatcaa
     1081 agcaaacttc aaaatccgtc ataacgtaga ggacggttct gttcagcttg ctgatcatta
     1141 tcagcaaaat acaccgatcg gtgacggccc ggttcttctt cctgataacc attatttatc
     1201 aactcaaagc gtattatcaa aagacccaaa tgaaaagcgt gaccacatgg tgctgcttga
     1261 atttgtgaca gctgctggta tcactcacgg catggatgag ctttataagt aagcatgcgc
     1321 cggtgattga ttgagcaagc tttatgcttg taaaccgttt tgtgaaaaaa tttttaaaat
     1381 aaaaaagggg acctctaggg tccccaatta attagtaata taatctatta aaggtcattc
     1441 aaaaggtcat ccaccggatc aattcccctg ctcgcgcagg ctgggtgcca agctctcggg
     1501 taacatcaag gcccgatcct tggagccctt gccctcccgc acgatgatcg tgccgtgatc
     1561 gaaatccaga tccttgaccc gcagttgcaa accctcactg atccggacga tacaaattcc
     1621 tcgtaggcgc tcgggacccc tatctagcga acttttagaa aagatataaa acatcagagt
     1681 atggacagtt gcggatgtac ttcagaaaag attagatgtc taaaaagctt gtagttaaag
     1741 ctttttagac atctaaatct aggtactaaa acaattcatc cagtaaaata taatatttta
     1801 ttttctccca atcaggcttg atccccagta agtcaaaaaa tagctcgaca tactgttctt
     1861 ccccgatatc ctccctgatc gaccggacgc agaaggcaat gtcataccac ttgtccgccc
     1921 tgccgcttct cccaagatca ataaagccac ttactttgcc atctttcaca aagatgttgc
     1981 tgtctcccag gtcgccgtgg gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa
     2041 aatcatacag ctcgcgcgga tctttaaatg gagtgtcttc ttcccagttt tcgcaatcca
     2101 catcggccag atcgttattc agtaagtaat ccaattcggc taagcggctg tctaagctat
     2161 tcgtataggg acaatccgat atgtcgatgg agtgaaagag cctgatgcac tccgcataca
     2221 gctcgataat cttttcaggg ctttgttcat cttcatactc ttccgagcaa aggacgccat
     2281 cggcctcact catgagcaga ttgctccagc catcatgccg ttcaaagtgc aggacctttg
     2341 gaacaggcag ctttccttcc agccatagca tcatgtcctt ttcccgttcc acatcatagg
     2401 tggtcccttt ataccggctg tccgtcattt ttaaatatag gttttcattt tctcccacca
     2461 gcttatatac cttagcagga gacattcctt ccgtatcttt tacgcagcgg tatttttcga
     2521 tcagtttttt caattccggt gatattctca ttttagccat ttattatttc cttcctcttt
     2581 tctacagtat ttaaagatac cccaagaagc taattataac aagacgaact ccaattcact
     2641 gttccttgca ttctaaaacc ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa
     2701 gttggcgtat aacatagtat cgacggagcc gattttgaaa ccacaattat gatagaattt
     2761 acaagctata aggttattgt cctgggtttc aagcattagt ccatgcaagt ttttatgctt
     2821 tgcccattct atagatatat tgataagcgc gctgcctatg ccttgccccc tgaaatcctt
     2881 acatacggcg atatcttcta tataaaagat atattatctt atcagtattg tcaatatatt
     2941 caaggcaatc tgcctcctca tcctcttcat cctcttcgtc ttggtagctt tttaaatatg
     3001 gcgcttcata gagtaattct gtaaaggtcc aattctcgtt ttcatacctc ggtataatct
     3061 tacctatcac ctcaaatggt tcgctgggtt tatcgtccgc gcagatcagt tggaagaatt
     3121 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc taacaattcg
     3181 ttcaagccga cgccgcttcg cggcgcggct taactcaagc gttagatgca ctaagcacat
     3241 aattgctcac agccaaacta tcaggtcaag tctgctttta ttatttttaa gcgtgcataa
     3301 taagccctac acaaattggg agatatatca tgaaaggctg gctttttctt gttatcgcaa
     3361 tagttggcga agtaatcgca acatccgcat taaaatctag cgagggcttt actaagctga
     3421 tccggtggat gaccttttga atgaccttta atagattata ttactaatta attggggacc
     3481 ctagaggtcc ccttttttat tttaaaaatt ttttcacaaa acggtttaca agcataaagc
     3541 ttgctcaatc aatcaccgga cgagcttgct caatcaatca ccggatcgat cctgttgcaa
     3601 taaacagaga tgctgcaacc ccccatgcaa taaaggcacc aatattaggc ataaccatgt
     3661 ttgataatga agttccgagt ttttgtaccc gaaccttcaa tgaagatttt cgttccatca
     3721 gaaatcccct cctttttatt ctaaaactac tgtaacattt ttcattttct aataaaactc
     3781 aaattttggc acaaaagttt tgccaaaatt tgttatccta agcggaagaa taggatttga
     3841 caaagcctga ctaagactga cttttttatc actaatattt ttctcatgtt aaaaacttgg
     3901 atatataact tttttaacag tttccttagg taatagagac gaaaaaaagc agcagcattc
     3961 ctcaagtttc cttttaaact tcaaactgaa tgtctcaact ctttcatata tttcctctga
     4021 tggtcaacat tcgttttatc acagcgaaag tggaatttgt tcttttatgt gactttacgg
     4081 ttgcttttag gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4141 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4201 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4261 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4321 gggcgctagc ggagtgtata tactggctta ctatgttggc actgatgagg gtgtcagtga
     4381 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     4441 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     4501 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     4561 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     4621 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     4681 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
     4741 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
     4801 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
     4861 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
     4921 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
     4981 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
     5041 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
     5101 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
     5161 aaaatatttc taggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat
     5221 cgcagtgatc cgcgatacaa attccccgta ggcgctaggg acctctttag ctccttggaa
     5281 gctgtcagta gtatacctaa taatttatct acattccctt tagtaacgtg taactttcca
     5341 aatttacaaa agcgactcat agaattattt cctcccgtta aataatagat aactattaaa
     5401 aatagacaat acttgctcat aagtaacggt acttaaattg tttactttgg cgtgtttcat
     5461 tgcttgatga aactgatttt tagtaaacag ttgacgatat tctcgattga cccattttga
     5521 aacaaagtac gtatatagct tccaatattt atctggaaca tctgtggtat ggcgggtaag
     5581 ttttattaag acactgttta cttttggttt aggatgaaag cattccgctg gcagcttaag
     5641 caattgctga atcgagactt gagtgtgcaa gagcaaccct agtgttcggt gaatatccaa
     5701 ggtacgcttg tagaatcctt cttcaacaat cagatagatg tcagacgcat ggctttcaaa
     5761 aaccactttt ttaataattt gtgtgcttaa atggtaagga atactcccaa caattttata
     5821 cctctgtttg ttagggaatt gaaactgtag aatatcttgg tgaattaaag tgacacgagt
     5881 attcagtttt aatttttctg acgataagtt gaatagatga ctgtctaatt caatagacgt
     5941 tacctgttta cttattttag ccagtttcgt cgttaaatgc cctttacctg ttccaatttc
     6001 gtaaacggta tcggtttctt ttaaattcaa ttgttttatt atttggttga gtactttttc
     6061 actcgttaaa aagttttgag aatattttat atttttgttc atgtaatcac tccttcttaa
     6121 ttacaaattt ttagcatcta atttaacttc aattcctatt atacaaaatt ttaagatact
     6181 gcactatcaa cacactctta agtttgcttc taagtcttat ttccataact tcttttacgt
     6241 ttccgccatt ctttgctgtt tcgattttta tgatatggtg caagtcagca cgaacacgaa
     6301 ccgtcttatc tcccattata tctttttttg cactgattgg tgtatcattt cgtttttctt
     6361 tttgtgcgcc taaatttccc acaatcactc acttctttct atttcttctt attcttattt
     6421 tatcatcaac aatcacaaat cacttgtgat ttgtgataag tgatttgtga ttaatatata
     6481 aaagccctct ttaaagggct tttatgttta ttttgagaaa gatataaaat caatatatcc
     6541 cttttctccg atttttacaa cggcattgta ggactttcta tctttcgttt tgattccttt
     6601 taccagggtt tcttttccct ctagtaattc ttttacattt gttttggtga gttttttctt
     6661 tctaaaatgt tcagctaaag taaacttaca ttcaggataa tttgaacaac cataaaacga
     6721 tttttttaat acaatattgt tgccacactt aggacatttt cctacaatac ttttttctgc
     6781 ttctttttct ttctgttcct ggtaatcaga aaaatttagt ttttctatat cgttaggtac
     6841 agcttccagt aaatgaacaa tgaatttttt gatattcgta ataaagttct cttgattgcc
     6901 ttctctttta ccgatttttt ttaaatacgt ttcccattta gccgtcattt cagcactcgt
     6961 taaaaggtgc tgactttcaa ctgcctggca caataatttt cctttttcag ttacaacaag
     7021 ctcggatcaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     7081 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     7141 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     7201 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     7261 gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac
     7321 caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg
     7381 cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca
     7441 gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac
     7501 ggcaaaagca ccgccggaca tcagcgccat tcgccattca ggctgcgcaa ctgttgggaa
     7561 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     7621 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//