LOCUS       sequence-211389-        6046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses high levels of gfp in mycobacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6046)
  AUTHORS   Eitson JL, Medeiros JJ, Hoover AR, Srivastava S, Roybal KT, Ainsa 
            JA, Hansen EJ, Gumbo T, van Oers NS
  TITLE     Mycobacterial shuttle vectors designed for high-level protein 
            expression in infected macrophages.
  JOURNAL   Appl Environ Microbiol. 2012 Oct;78(19):6829-37. doi: 
            10.1128/AEM.01674-12. Epub 2012 Jul 20.
  PUBMED    22820329
REFERENCE   2  (bases 1 to 6046)
  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: 
            "10.1128/AEM.01674-12"; journalName: "Appl Environ Microbiol"; date:
            "2012-10"; volume: "78"; issue: "19"; pages: "6829-37"
COMMENT     Sequence Label: pSUM-Kan-MCS1-gfp
FEATURES             Location/Qualifiers
     source          1..6046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             83..796
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(113..141)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     736..756
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    800..856
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1204..1221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1338..1883)
                     /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(1418..1437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4856..5650
                     /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(4910..4929)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5520..5539
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6015..6037
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6029..6046
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6030..6046
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gccaagcttg ggctgcaggt cgactctaga ggatccccgg gcgagctcga attctgatta
       61 actttataag gaggaaaaac atatgagtaa aggagaagaa cttttcactg gagttgtccc
      121 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
      181 tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact
      241 acctgttcca tggccaacac ttgtcactac tttgacttat ggtgttcaat gcttttcaag
      301 atacccagat catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt
      361 acaggaaaga actatatttt tcaaagatga cgggaactat aagacacgtg ctgaagtcaa
      421 gtttgaaggt gatacacttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga
      481 tggaaacatt cttggacaca aattggaata caactataac tcacacaatg tatacatcat
      541 ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa attagacaca acattgaaga
      601 tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg atggccctgt
      661 ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag atcccaacga
      721 aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat
      781 ggatgaacta tacaaatagg aattcgagct cggtacccgg ggatcctcta gagtcgacct
      841 gcaggcatgc aagcttggct gcaggtcgac ggatccgatt ggagccctcc actatttgga
      901 tcagagcccg ggacgctcgc ccggaccgga gttggcggtc ggtccggggc gtgccccgga
      961 accgtccgaa ttaccaggtg attcggcatt cgtccgcgcc ctcgcgccgt cgtcgcgggt
     1021 gccgacgcag gggttagccg attgccatct agcactctat acatgagagt gctagcggag
     1081 tgtatatact ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg
     1141 caggagaaaa aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc
     1201 ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta
     1261 cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg
     1321 gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga
     1381 cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1441 ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg
     1501 ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc
     1561 caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa
     1621 ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg
     1681 taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga
     1741 caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc
     1801 agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta
     1861 cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa tatcgagccg
     1921 agaacgttat cgaagttggt catgtgtaat cccctcgttt gaactttgga ttaagcgtag
     1981 atacaccctt ggacaagcca gttggattcg gagacaagca aattcagcct taaaaagggc
     2041 gaggccctgc ggtggtggaa caccgcaggg cctctaaccg ctcgacgcgc tgcaccaacc
     2101 agcccgcgaa cggctggcag ccagcgtaag gcgcggctca tcgggcggcg ttcgccacga
     2161 tgtcctgcac ttcgagccaa gcctcgaaca cctgctggtg tgcacgactc acccggttgt
     2221 tgacaccgcg cgcggccgtg cgggctcggt ggggcggctc tgtcgccctt gccagcgtga
     2281 gtagcgcgta cctcacctcg cccaacaggt cgcacacagc cgattcgtac gccataaagc
     2341 caggtgagcc caccagctcc gtaagttcgg gcgctgtgtg gctcgtaccc gcgcattcag
     2401 gcggcagggg gtctaacggg tctaaggcgg cgtgtacggc cgccacagcg gctctcagcg
     2461 gcccggaaac gtcctcgaaa cgacgcatgt gttcctcctg gttggtacag gtggttgggg
     2521 gtgctcggct gtcgctggtg ttccaccacc agggctcgac gggagagcgg gggagtgtgc
     2581 agttgtgggg tggcccctca gcgaaatatc tgacttggag ctcgtgtcgg accatacacc
     2641 ggtgattaat cgtggtctac taccaagcgt gagccacgtc gccgacgaat ttgagcagct
     2701 ctggctgccg tactggccgc tggcaagcga cgatctgctc gaggggatct accgccaaag
     2761 ccgcgcgtcg gccctaggcc gccggtacat cgaggcgaac ccaacagcgc tggcaaacct
     2821 gctggtcgtg gacgtagacc atccagacgc agcgctccga gcgctcagcg cccgggggtc
     2881 ccatccgctg cccaacgcga tcgtgggcaa tcgcgccaac ggccacgcac acgcagtgtg
     2941 ggcactcaac gcccctgttc cacgcaccga atacgcgcgg cgtaagccgc tcgcatacat
     3001 ggcggcgtgc gccgaaggcc ttcggcgcgc cgtcgacggc gaccgcagtt actcaggcct
     3061 catgaccaaa aaccccggcc acatcgcctg ggaaacggaa tggctccact cagatctcta
     3121 cacactcagc cacatcgagg ccgagctcgg cgcgaacatg ccaccgccgc gctggcgtca
     3181 gcagaccacg tacaaagcgg ctccgacgcc gctagggcgg aattgcgcac tgttcgattc
     3241 cgtcaggttg tgggcctatc gtcccgccct catgcggatc tacctgccga cccggaacgt
     3301 ggacggactc ggccgcgcga tctatgccga gtgccacgcg cgaaacgccg aattcccgtg
     3361 caacgacgtg tgtcccggac cgctaccgga cagcgaggtc cgcgccatcg ccaacagcat
     3421 ttggcgttgg atcacaacca agtcgcgcat ttgggcggac gggatcgtgg tctacgaggc
     3481 cacactcagt gcgcgccagt cggccatctc gcggaagggc gcagcagcgc gcacggcggc
     3541 gagcacagtt gcgcggcgcg caaagtccgc gtcagccatg gaggcattgc tatgagcgac
     3601 ggctacagcg acggctacaa ccggcagccg actgtccgca aaaagcggcg cgtgaccgcc
     3661 gccgaaggcg ctcgaatcac cggactatcc gaacgccacg tcgtccggct cgtggcgcag
     3721 gaacgcagcg agtggctcgc cgagcaggct gcacgccgcg aacgcatccg cgcctatcac
     3781 gacgacgagg gccactcttg gccgcaaacg gccaaacatt tcgggctgca tctggacacc
     3841 gttaagcgac tcggctatcg ggcgaggaaa gagcgtgcgg cagaacagga agcggctcaa
     3901 aaggcccaca acgaagccga caatccaccg ctgttctaac gcaattgggg agcgggtgtc
     3961 gcgggggttc cgtggggggt tccgttgcaa cgggtcggac aggtaaaagt cctggtagac
     4021 gctagttttc tggtttgggc catgcctgtc tcgttgcgtg tttcgttgcg cccgttttga
     4081 ataccagcca gacgagacgg ggttctacga atcttggtcg ataccaagcc atttccgctg
     4141 aatatcgggg agctcaccgc cagaatcggt ggttgtggtg atgtacgtgg cgaactccgt
     4201 tgtagtgcct gtggtggcat ccgtggccac tctcgttgca cggttcgttg tgccgttaca
     4261 ggccccgttg acagctcacc gaacgtagtt aaaacatgct ggtcaaacta ggtttaccaa
     4321 cgatacgagt cagctcatct agggccagtt ctaggcgttg ttcgttgcgc ggttcgttgc
     4381 gcatgtttcg tgtggttgct agatggctcc gcaaccacac gcttcgaggt tgagtgcttc
     4441 cagcacgggc gcgatccaga agaacttcgt cgtgcgactg tcctcgttat ttctaggcac
     4501 agcttcacgc tgccgcaagc actcagggcg caagggctgc taaaggaagc ggaacacgta
     4561 gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg
     4621 gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg
     4681 atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc
     4741 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
     4801 ctgatggcgc aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgcatgat
     4861 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4921 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4981 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactccaaga
     5041 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5101 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5161 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5221 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5281 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5341 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcggatgc ccgacggcga
     5401 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5461 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5521 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5581 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5641 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5701 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5761 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5821 cccctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg
     5881 acatcagcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5941 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     6001 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagt
//
