LOCUS       sequence-370623-        6440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for integrative expression, pUC19 backbone, Apr resistant 
            casette, pCMP1::eGFP expression cassette, gateway compatible.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6440)
  AUTHORS   Stevens DM, Tang A, Coaker G
  TITLE     A Genetic Toolkit for Investigating Clavibacter Species: Markerless 
            Deletion, Permissive Site Identification, and an Integrative 
            Plasmid.
  JOURNAL   Mol Plant Microbe Interact. 2021 Dec;34(12):1336-1345. doi: 
            10.1094/MPMI-07-21-0171-TA. Epub 2021 Dec 10.
  PUBMED    34890250
REFERENCE   2  (bases 1 to 6440)
  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"; doi: 
            "10.1094/MPMI-07-21-0171-TA"; journalName: "Mol Plant Microbe 
            Interact"; date: "2021-12"; volume: "34"; issue: "12"; pages: 
            "1336-1345"
COMMENT     Sequence Label: pSelAct-Express
FEATURES             Location/Qualifiers
     source          1..6440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     170..189
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     429..450
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     oriT            1348..1457
                     /note="incP origin of transfer"
     CDS             1773..2573
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     rep_origin      2690..3278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3179..3198
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3432..3449
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3566..3587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3602..3607,3608..3625,3626..3632)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3640..3656
                     /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)."
     primer_bind     3645..3667
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3664..3680
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3664..3680
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3699..3719
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3701..3719
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3749..3765
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3800..3924
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(3965..4270)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(4067..4086)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(4590..5270)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     5185..5204
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(5324..5330,5331..5348,5349..5354))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(5379..5503)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5513..5529)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5513..5529)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5561..5580)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5562..5580)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5590..5607)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5590..5606)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5599..5621)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5748..6176
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5835..5854)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6045..6066
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(6216..6440,1..489,490..492,493..495))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVYGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6261..6282)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
       61 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
      121 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
      181 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
      241 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
      301 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
      361 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccgtacacgc tgaacttgtg
      421 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
      481 gcccttgctc accatggtag ttctcctgag gacgtcttgt caccattttt aacgattgcg
      541 taatcgcgca aagttccaac aacctgagcc gaattagcat taatcgtgat gaccttaccg
      601 atcatcgccg ggtcgaacgc ggtgtacgtc ttactcatgc tcaatcatct ccataagcgg
      661 ctcaagctct tcggcaagtt cgggtcgatc cataagaatc tccatataaa cgtcctcaaa
      721 aacagaatcg ttttgaaaag tcaggcgtcg tgcagagtag cccgtcctga tgcatcagta
      781 cgccggtgat ctcagctgta ggaaagagaa gaaggttagt agtcgacggt atcgataagc
      841 ttgatgatat catgcgaaac gatcctcatc ctgtctcttg atcagatcga tcttgatccc
      901 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
      961 aaccttacca gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc
     1021 ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
     1081 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     1141 cggactggct ttctacgtgt tccgcttcct ttagcagccc ttgcgccctg agtgcttgcg
     1201 gcagcgtgaa gctatcgaat tattaacgct tacaatttag gtggcacttt tcggggaaat
     1261 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1321 cctagaaagt ataggaactt cgaagttccc gccagcctcg cagagcagga ttcccgttga
     1381 gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta
     1441 cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc
     1501 tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa
     1561 tgaccccgaa gcagggttat gcagcggaaa atgcagctca cggtaactga tgccgtattt
     1621 gcagtaccag cgtacggccc acagaatgat gtcacgctga aaatgccggc ctttgaatgg
     1681 gttcatgtgc agctccataa gcaaaagggg atgataagtt tatcaccacc gactatttgc
     1741 aacagtgccg ttgatcgtgc tatgatcgac tgatgtcatc agcggtggag tgcaatgtcg
     1801 tgcaatacga atggcgaaaa gccgagctca tcggtcagct tctcaacctt ggggttaccc
     1861 ccggcggtgt gctgctggtc cacagctcct tccgtagcgt ccggcccctc gaagatgggc
     1921 cacttggact gatcgaggcc ctgcgtgctg cgctgggtcc gggagggacg ctcgtcatgc
     1981 cctcgtggtc aggtctggac gacgagccgt tcgatcctgc cacgtcgccc gttacaccgg
     2041 accttggagt tgtctctgac acattctggc gcctgccaaa tgtaaagcgc agcgcccatc
     2101 catttgcctt tgcggcagcg gggccacagg cagagcagat catctctgat ccattgcccc
     2161 tgccacctca ctcgcctgca agcccggtcg cccgtgtcca tgaactcgat gggcaggtac
     2221 ttctcctcgg cgtgggacac gatgccaaca cgacgctgca tcttgccgag ttgatggcaa
     2281 aggttcccta tggggtgccg agacactgca ccattcttca ggatggcaag ttggtacgcg
     2341 tcgattatct cgagaatgac cactgctgtg agcgctttgc cttggcggac aggtggctca
     2401 aggagaagag ccttcagaag gaaggtccag tcggtcatgc ctttgctcgg ttgatccgct
     2461 cccgcgacat tgtggcgaca gccctgggtc aactgggccg agatccgttg atcttcctgc
     2521 atccgccaga ggcgggatgc gaagaatgcg atgccgctcg ccagtcgatt ggctgagctc
     2581 ataagttcct attccgaagt tcctattctc tagtcatgac caaaatccct taacgtgagt
     2641 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2701 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2761 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2821 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2881 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2941 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3001 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3061 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3121 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3181 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3241 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3301 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3361 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3421 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     3481 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     3541 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     3601 ctttacactt tatgctcccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     3661 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
     3721 aacaaaagct gggtaccggg ccccccctcg aggtcgacgg tatcgataag cttgatatcg
     3781 aattcctgca gcccatcgaa ccactttgta caagaaagct gaacgagaaa cgtaaaatga
     3841 tataaatatc aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa
     3901 aacacaacat atccagtcac tatggtcgac ctgcagactg gctgtgtata agggagcctg
     3961 acatttatat tccccagaac atcaggttaa tggcgttttt gatgtcattt tcgcggtggc
     4021 tgagatcagc cacttcttcc ccgataacgg agaccggcac actggccata tcggtggtca
     4081 tcatgcgcca gctttcatcc ccgatatgca ccaccgggta aagttcacgg gggactttat
     4141 ctgacagcag acgtgcactg gccaggggga tcaccatccg tcgcccgggc gtgtcaataa
     4201 tatcactctg tacatccaca aacagacgat aacggctctc tcttttatag gtgtaaacct
     4261 taaactgcat ttcaccagcc cctgttctcg tcagcaaaag agccgttcat ttcaataaac
     4321 cgggcgacct cagccatccc ttcctgattt tccgctttcc agcgttcggc acgcagacga
     4381 cgggcttcat tctgcatggt tgtgcttacc agaccggaga tattgacatc atatatgcct
     4441 tgagcaactg atagctgtcg ctgtcaactg tcactgtaat acgctgcttc atagcatacc
     4501 tctttttgac atacttcggg tatacatatc agtatatatt cttataccgc aaaaatcagc
     4561 gcgcaaatac gcatactgtt atctggcttt tagtaagccg gatcctctag attacgcccc
     4621 gcctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     4681 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     4741 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     4801 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
     4861 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     4921 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     4981 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     5041 attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     5101 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     5161 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     5221 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     5281 ttagctcctg aaaatctcga cggatcctaa ctcaaaatcc acacattata cgagccggaa
     5341 gcataaagtg taaagcctgg ggtgcctaat gcggccgcca tagtgactgg atatgttgtg
     5401 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
     5461 tatcatttta cgtttctcgt tcagcttttt tgtacaaact tgtttgatgg gggatccact
     5521 agttctagag cggccgccac cgcggtggag ctccaattcg ccctatagtg agtcgtatta
     5581 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     5641 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     5701 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     5761 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5821 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5881 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5941 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     6001 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     6061 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     6121 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     6181 caaaatctag cgaatttcga caagcacagc caagcttact tgtacagctc gtccatgccg
     6241 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     6301 gggtctttgc tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg
     6361 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg
     6421 atgttgtggc ggatcttgaa
//
