LOCUS       sequence-211324-        6082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses gfp in mycobacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6082)
  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 6082)
  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-GroEL-gfp
FEATURES             Location/Qualifiers
     source          1..6082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     88..105
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             623..1333
                     /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="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(650..678)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1273..1293
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1345..1362)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1345..1361)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1354..1376)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1741..2535)
                     /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(1852..1871)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2462..2481
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      5508..6053
                     /direction=RIGHT
                     /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     5954..5973
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc
       61 gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc
      121 ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca
      181 cttcactgac accctcatca gtgccaacat agtaagccag tatatacact ccgctagatg
      241 catgctcgag cggccgccag tgtgatggat atctgcagaa ttcgcccttc tagctagcgg
      301 caggtgcggt gacccggaca ttgcacctgg cgtacccgga cacgctggca accaggaagc
      361 aagggggcgc ccttgagtgc tagcactctc atgtatagag tgctagatgg caatcggcta
      421 acccctgcgt cggcacccgc gacgacggcg cgagggcgcg gacgaatgcc gaatcacctg
      481 gtaattcgga cggttccggg gcacgccccg gaccgaccgc caactccggt ccgggcgagc
      541 gtcccgggct ctgatccaaa tagtggaggg ctccaatcgg atccattaac tttataagga
      601 ggaaaaacat atgaaagctt tcagtaaagg agaagaactt ttcactggag ttgtcccaat
      661 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
      721 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
      781 tgttccatgg ccaacacttg tcactacttt gacttatggt gttcaatgct tttcaagata
      841 cccagatcat atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca
      901 ggaaagaact atatttttca aagatgacgg gaactataag acacgtgctg aagtcaagtt
      961 tgaaggtgat acacttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg
     1021 aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc
     1081 agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg
     1141 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     1201 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa
     1261 gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
     1321 tgaactatac aaatgaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     1381 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     1441 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     1501 gcgctgatgt ccggcggtgc ttttgccgtt acgcaccacc ccgtcagtag ctgaacagga
     1561 gggacagggg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     1621 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     1681 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     1741 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     1801 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     1861 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     1921 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     1981 gacgagatcc tcgccgtcgg gcatccgcgc cttgagcctg gcgaacagtt cggctggcgc
     2041 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     2101 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     2161 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     2221 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     2281 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     2341 cgctgcctcg tcttggagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     2401 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     2461 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     2521 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct
     2581 gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac
     2641 cttaccagag ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca
     2701 gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt
     2761 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg
     2821 actggctttc tacgtgttcc gcttccttta gcagcccttg cgccctgagt gcttgcggca
     2881 gcgtgaagct gtgcctagaa ataacgagga cagtcgcacg acgaagttct tctggatcgc
     2941 gcccgtgctg gaagcactca acctcgaagc gtgtggttgc ggagccatct agcaaccaca
     3001 cgaaacatgc gcaacgaacc gcgcaacgaa caacgcctag aactggccct agatgagctg
     3061 actcgtatcg ttggtaaacc tagtttgacc agcatgtttt aactacgttc ggtgagctgt
     3121 caacggggcc tgtaacggca caacgaaccg tgcaacgaga gtggccacgg atgccaccac
     3181 aggcactaca acggagttcg ccacgtacat caccacaacc accgattctg gcggtgagct
     3241 ccccgatatt cagcggaaat ggcttggtat cgaccaagat tcgtagaacc ccgtctcgtc
     3301 tggctggtat tcaaaacggg cgcaacgaaa cacgcaacga gacaggcatg gcccaaacca
     3361 gaaaactagc gtctaccagg acttttacct gtccgacccg ttgcaacgga accccccacg
     3421 gaacccccgc gacacccgct ccccaattgc gttagaacag cggtggattg tcggcttcgt
     3481 tgtgggcctt ttgagccgct tcctgttctg ccgcacgctc tttcctcgcc cgatagccga
     3541 gtcgcttaac ggtgtccaga tgcagcccga aatgtttggc cgtttgcggc caagagtggc
     3601 cctcgtcgtc gtgataggcg cggatgcgtt cgcggcgtgc agcctgctcg gcgagccact
     3661 cgctgcgttc ctgcgccacg agccggacga cgtggcgttc ggatagtccg gtgattcgag
     3721 cgccttcggc ggcggtcacg cgccgctttt tgcggacagt cggctgccgg ttgtagccgt
     3781 cgctgtagcc gtcgctcata gcaatgcctc catggctgac gcggactttg cgcgccgcgc
     3841 aactgtgctc gccgccgtgc gcgctgctgc gcccttccgc gagatggccg actggcgcgc
     3901 actgagtgtg gcctcgtaga ccacgatccc gtccgcccaa atgcgcgact tggttgtgat
     3961 ccaacgccaa atgctgttgg cgatggcgcg gacctcgctg tccggtagcg gtccgggaca
     4021 cacgtcgttg cacgggaatt cggcgtttcg cgcgtggcac tcggcataga tcgcgcggcc
     4081 gagtccgtcc acgttccggg tcggcaggta gatccgcatg agggcgggac gataggccca
     4141 caacctgacg gaatcgaaca gtgcgcaatt ccgccctagc ggcgtcggag ccgctttgta
     4201 cgtggtctgc tgacgccagc gcggcggtgg catgttcgcg ccgagctcgg cctcgatgtg
     4261 gctgagtgtg tagagatctg agtggagcca ttccgtttcc caggcgatgt ggccggggtt
     4321 tttggtcatg aggcctgagt aactgcggtc gccgtcgacg gcgcgccgaa ggccttcggc
     4381 gcacgccgcc atgtatgcga gcggcttacg ccgcgcgtat tcggtgcgtg gaacaggggc
     4441 gttgagtgcc cacactgcgt gtgcgtggcc gttggcgcga ttgcccacga tcgcgttggg
     4501 cagcggatgg gacccccggg cgctgagcgc tcggagcgct gcgtctggat ggtctacgtc
     4561 cacgaccagc aggtttgcca gcgctgttgg gttcgcctcg atgtaccggc ggcctagggc
     4621 cgacgcgcgg ctttggcggt agatcccctc gagcagatcg tcgcttgcca gcggccagta
     4681 cggcagccag agctgctcaa attcgtcggc gacgtggctc acgcttggta gtagaccacg
     4741 attaatcacc ggtgtatggt ccgacacgag ctccaagtca gatatttcgc tgaggggcca
     4801 ccccacaact gcacactccc ccgctctccc gtcgagccct ggtggtggaa caccagcgac
     4861 agccgagcac ccccaaccac ctgtaccaac caggaggaac acatgcgtcg tttcgaggac
     4921 gtttccgggc cgctgagagc cgctgtggcg gccgtacacg ccgccttaga cccgttagac
     4981 cccctgccgc ctgaatgcgc gggtacgagc cacacagcgc ccgaacttac ggagctggtg
     5041 ggctcacctg gctttatggc gtacgaatcg gctgtgtgcg acctgttggg cgaggtgagg
     5101 tacgcgctac tcacgctggc aagggcgaca gagccgcccc accgagcccg cacggccgcg
     5161 cgcggtgtca acaaccgggt gagtcgtgca caccagcagg tgttcgaggc ttggctcgaa
     5221 gtgcaggaca tcgtggcgaa cgccgcccga tgagccgcgc cttacgctgg ctgccagccg
     5281 ttcgcgggct ggttggtgca gcgcgtcgag cggttagagg ccctgcggtg ttccaccacc
     5341 gcagggcctc gcccttttta aggctgaatt tgcttgtctc cgaatccaac tggcttgtcc
     5401 aagggtgtat ctacgcttaa tccaaagttc aaacgagggg attacacatg accaacttcg
     5461 ataacgttct cggctcgata ttttatctga ttaataagat gatcttcttg agatcgtttt
     5521 ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg
     5581 aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca
     5641 ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct
     5701 aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc
     5761 aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca
     5821 gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac
     5881 gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga
     5941 gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact
     6001 gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct
     6061 ttgccgcggc cctctcactt cc
//
