LOCUS       sequence-11493-e        6778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6778)
  AUTHORS   Blokpoel MC, Murphy HN, O'Toole R, Wiles S, Runn ES, Stewart GR, 
            Young DB, Robertson BD
  TITLE     Tetracycline-inducible gene regulation in mycobacteria.
  JOURNAL   Nucleic Acids Res. 2005  . 33(2):e22.
  PUBMED    15687380
REFERENCE   2  (bases 1 to 6778)
  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: "Nucleic 
            Acids Res. 2005  . 33(2):e22."
COMMENT     Sequence Label: pMIND
FEATURES             Location/Qualifiers
     source          1..6778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..35
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     19..35
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1916..1933)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1916..1932)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1925..1947)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4745..5539
                     /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(4799..4818)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5409..5428
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      5823..6411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6312..6331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6565..6582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6699..6720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6735..6740,6741..6758,6759..6765)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(6773..6778,1..11)
                     /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     join(6778,1..22)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattag cccgcctaat
       61 gagcgggctt ttttttgaat tagcccgcct aatgagcggg cttttttttg aattcgagct
      121 cggtacctct agatcacgat tcgctcgagg tcaatgacgc tgtgcgggag tgagagaatc
      181 cgtcgatcag tgctgtcagg ccgaaatcga attcccggtc gcgttcagga acgctcgcat
      241 tgccgacctc tgcggtttgc tcctccagta cgacaccgac ggtgtatcgg ctgaccgcga
      301 tgagcatgct gatggctgtc tgttcatcga atccctgaga cacgagaagc ccgacttgct
      361 gaccggaagc ctcagctcct tcgtcggtgt cgtcgccgga tggtctgcgg tggtgctcag
      421 cgtggagacg cgcaccgtca cggaccgcgt gtagtgcgtg gcggaaactt cttgcgtttc
      481 gcaagagaaa tgcgtcccat ttctcgtcgg actcggggaa ggaagcgtga tgctctcggt
      541 caagcacgtc gctcgccagc gctgcgagga gttcggcctt cgtgcggaag tgccagtaga
      601 ggccgggctg ctgtacctgt aagtgagccg ccagcgcgcg agtggtgaag ccatcgagcc
      661 cagtctcgtc gagcacctgc cgggccccga gcaacacgga cgtgcggtcg agacgcttcc
      721 ggtggtgagt catagttgca ctttatcatc gataacttta tcttagataa agtgactgct
      781 cgctactctc atctgactgc tcgctactct catcgtggaa tcctgacagg atcccttaag
      841 aactgcagtc catatggata tccttaatta agtatgcatc gacgcgttgt acaggactag
      901 tgcatgcgtg acacaagaat ccctgttact tctcgaccgt attgattcgg atgattccta
      961 cgcgagcctg cggaacgacc aggaattctg ggagccgctg gcccgccgag ccctggagga
     1021 gctcgggctg ccggtgccgc cggtgctgcg ggtgcccggc gagagcacca accccgtact
     1081 ggtcggcgag cccggcccgg tgatcaagct gttcggcgag cactggtgcg gtccggagag
     1141 cctcgcgtcg gagtcggagg cgtacgcggt cctggcggac gccccggtgc cggtgccccg
     1201 cctcctcggc cgcggcgagc tgcggcccgg caccggagcc tggccgtggc cctacctggt
     1261 gatgagccgg atgaccggca ccacctggcg gtccgcgatg gacggcacga ccgaccggaa
     1321 cgcgctgctc gccctggccc gcgaactcgg ccgggtgctc ggccggctgc acagggtgcc
     1381 gctgaccggg aacaccgtgc tcacccccca ttccgaggtc ttcccggaac tgctgcggga
     1441 acgccgcgcg gcgaccgtcg aggaccaccg cgggtggggc tacctctcgc cccggctgct
     1501 ggaccgcctg gaggactggc tgccggacgt ggacacgctg ctggccggcc gcgaaccccg
     1561 gttcgtccac ggcgacctgc acgggaccaa catcttcgtg gacctggccg cgaccgaggt
     1621 caccgggatc gtcgacttca ccgacgtcta tgcgggagac tcccgctaca gcctggtgca
     1681 actgcatctc aacgccttcc ggggcgaccg cgagatcctg gccgcgctgc tcgacggggc
     1741 gcagtggaag cggaccgagg acttcgcccg cgaactgctc gccttcacct tcctgcacga
     1801 cttcgaggtg ttcgaggaga ccccgctgga tctctccggc ttcaccgatc cggaggaact
     1861 ggcgcagttc ctctgggggc cgccagacac cgcccccggc gcctaaaagc ttggcactgg
     1921 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     1981 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2041 cccaacagtt gcgcagcctg aatggcgaat ggcgataagc tagcccaacc agcccgcgaa
     2101 cggctggcag ccagcgtaag gcgcggctca tcgggcggcg ttcgccacga tgtcctgcac
     2161 ttcgagccaa gcctcgaaca cctgctggtg tgcacgactc acccggttgt tgacaccgcg
     2221 cgcggccgtg cgggctcggt ggggcggctc tgtcgccctt gccagcgtga gtagcgcgta
     2281 cctcacctcg cccaacaggt cgcacacagc cgattcgtac gccataaagc caggtgagcc
     2341 caccagctcc gtaagttcgg gcgctgtgtg gctcgtaccc gcgcattcag gcggcagggg
     2401 gtctaacggg tctaaggcgg cgtgtacggc cgccacagcg gctctcagcg gcccggaaac
     2461 gtccttgaaa cgacgtatgt gttcctcctg gttggtacag gtggttgggg gtgctcggct
     2521 gtcgctggtg ttccaccacc agggctcgac gggagagcgg gggagtgtgc agttgtgggg
     2581 tggcccctca gcgaaatatc tgacttggag ctcgtgtcgg accatacacc ggtgattaat
     2641 cgtggtctac taccaagcgt gagccacgtc gccgacgaat ttgagcagct ctggctgccg
     2701 tactggccgc tggcaagcga cgatctgctc gaggggatct accgccaaag ccgcgcgtcg
     2761 gccctaggcc gccggtacat cgaggcgaac ccaacagcgc tggcaaacct gctggtcgtg
     2821 gacgtagacc atccagacgc agcgctccga gcgctcagcg cccgggggtc ccatccgctg
     2881 cccaacgcga tcgtgggcaa tcgcgccaac ggccacgcac acgcagtgtg ggcactcaac
     2941 gcccctgttc cacgcaccga atacgcgcgg cgtaagccgc tcgcatacat ggcggcgtgc
     3001 gccgaaggcc ttcggcgcgc cgtcgacggc gaccgcagtt actcaggcct catgaccaaa
     3061 aaccccggcc acatcgcctg ggaaacggaa tggctccact cagatctcta cacactcagc
     3121 cacatcgagg ccgagctcgg cgcgaacatg ccaccgccgc gctggcgcca gcagaccacg
     3181 tacaaagcgg ctccgacgcc gctagggcgg aattgcgcac tgttcgattc cgtcaggttg
     3241 tgggcctatc gtcccgccct catgcggatc tacctgccga cccggaacgt ggacggactc
     3301 ggccgcgcga tctatgccga gtgccacgtc cgaaacgccg agttcccgtg caacgacgtg
     3361 tgtcccggac cgctaccgga cagcgaggtc cgcgccatcg ccaacagcat ttggcgttgg
     3421 atcacaacca agtcgcgcat ttgggcggac gggatcgtgg tctacgaggc cacactcagt
     3481 gcgcgccagt cggccatctc gcggaagggc gcagcagcgc gcacggcggc gagcacagtt
     3541 gcgcggcgcg caaagtccgc gtcagccatg gaggcattgc tatgagcgac ggctacagcg
     3601 acggctacaa ccggcagccg actgtccgca aaaagcggcg cgtgaccgcc gccgaaggcg
     3661 ctcgaatcac cggactatcc gaacgccacg tcgtccggct cgtggcgcag gaacgcagcg
     3721 agtggctcgc cgagcaggct gcacgccgcg aacgcatccg cgcctatcac gacgacgagg
     3781 gccactcttg gccgcaaacg gccaaacatt tcgggctgca tctggacacc gttaagcgac
     3841 tcggctatcg ggcgaggaaa gagcgtgcgg cagaacagga agcggctcaa aaggcccaca
     3901 acgaagccga caatccaccg ctgttctaac gcaattgggg agcgggtgtc gcgggggttc
     3961 cgtggggggt tccgttgcaa cgggtcggac aggtaaaagt cctggtagac gctagttttc
     4021 tggtttgggc catgcctgtc tcgttgcgtg tttcgttgcg cccgttttga ataccagcca
     4081 gacgagacgg ggttctacga atcttggtcg ataccaagcc atttccgctg aatatcgggg
     4141 agctcaccgc cagaatcggt ggttgtggtg atgtacgtgg cgaactccgt tgtagtgcct
     4201 gtggtggcat ccgtggccac tctcgttgca cggttcgttg tgccgttaca ggccccgttg
     4261 acagctcacc gaacgtagtt aaaacatgct ggtcaaacta ggtttaccaa cgatacgagt
     4321 cagctcatct agggccagtt ctaggcgttg ttcgttgcgc ggttcgttgc gcatgtttcg
     4381 tgtggttgct agcttcacgc tgccgcaagc actcagggcg caagggctgc taaaggaagc
     4441 ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact
     4501 gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc
     4561 ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag
     4621 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     4681 cgccaaggat ctgatggcgc agggatcaag atctgatcaa gagacaggat gaggatcgtt
     4741 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4801 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4861 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4921 actccaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4981 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5041 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5101 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5161 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5221 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcggatgcc
     5281 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5341 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5401 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5461 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5521 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5581 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5641 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5701 ttcgcccacc ccaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5761 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5821 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5881 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5941 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6001 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6061 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6121 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6181 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6241 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6301 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6361 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6421 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6481 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6541 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6601 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     6661 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     6721 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtga
//
