LOCUS       sequence-11542-e        8798 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 8798)
  AUTHORS   Williams KJ, Joyce G, Robertson BD
  TITLE     Improved mycobacterial tetracycline inducible vectors.
  JOURNAL   Plasmid. 2010 Sep;64(2):69-73. doi: 10.1016/j.plasmid.2010.04.003. 
            Epub 2010 Apr 29.
  PUBMED    20434484
REFERENCE   2  (bases 1 to 8798)
  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: 
            "10.1016/j.plasmid.2010.04.003"; journalName: "Plasmid"; date: 
            "2010-09"; volume: "64"; issue: "2"; pages: "69-73"
COMMENT     Sequence Label: pMEND-Lx
FEATURES             Location/Qualifiers
     source          1..8798
                     /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(3936..3953)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3936..3952)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3945..3967)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             6765..7559
                     /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(6819..6838)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7429..7448
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      7843..8431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8332..8351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8585..8602
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8719..8740
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8755..8760,8761..8778,8779..8785)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(8793..8798,1..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(8798,1..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 gggtgggctg 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 atccggagga
      841 aatgttatga aatttggaaa cttccttctc acttatcagc cacctgagct atctcagacc
      901 gaagtgatga agcgattggt taatctgggc aaagcgtctg aaggttgtgg cttcgacacc
      961 gtttggttgc tagagcacca cttcactgaa tttgggttgt tagggaatcc ttatgttgct
     1021 gccgcacacc tattaggtgc gacagaaacg ctcaacgttg gcactgcagc tatcgtattg
     1081 ccgactgccc atccggttcg acaagcagaa gacgtaaacc tactggatca aatgtcaaaa
     1141 ggacgattcc gttttggtat ttgtcgcggt ttgtacgata aagattttcg tgtctttggt
     1201 acagacatgg ataacagccg agccttaatg gactgttggt atgacttgat gaaagaaggc
     1261 ttcaatgaag gctatatcgc ggcggataac gaacatatta agttcccgaa aatccaactg
     1321 aatccatcgg cttacacaca aggtggtgct cctgtttatg tcgtcgcgga gtcagcatca
     1381 acgacagaat gggctgcaga gcgtggccta ccaatgattc taagctggat catcaacact
     1441 cacgagaaga aagcgcagct tgatctttac aacgaagtcg cgactgaaca tggctacgat
     1501 gtgactaaga ttgaccactg tttgtcttac atcacctccg tcgatcatga ctcaaataga
     1561 gccaaagata tttgccgcaa cttcttgggc cattggtacg actcatacgt gaatgccacc
     1621 aagatttttg acgactctga ccaaacaaaa ggttacgact tcaataaagg tcaatggcgt
     1681 gattttgtgt tgaaaggcca caaagacacc aatcgccgaa ttgattacag ctacgaaatc
     1741 aacccagtag ggacgcctga agagtgtatc gcgattatcc agcaagatat tgatgcgacg
     1801 ggtattgaca atatttgttg tggttttgaa gcaaacggtt ctgaagaaga aattatcgca
     1861 tctatgaagc tattccagtc tgatgtgatg ccatatctca aagaaaaaca gtaattaata
     1921 ttttctaaaa ggaaagagac atgaaatttg gattattctt cctcaatttt atgaactcaa
     1981 agcgttcttc tgatcaagtc atcgaagaaa tgttagatac cgcacattac gtagatcagt
     2041 tgaagtttga cacgttggct gtttacgaaa accatttctc gaacaatggt gtggttggtg
     2101 ccccactaac agtggctggt tttttacttg gtatgacaaa gaacgccaaa gtggcttcgt
     2161 tgaatcacgt cattaccacg catcatccag tacgtgtggc ggaagaagcg tgtctacttg
     2221 accaaatgag tgaaggccgt tttgcctttg gctttagtga ttgtgaaaag agtgcagata
     2281 tgcgcttctt taatcgacca acggattctc agtttcagtt gttcagtgag tgtcacaaga
     2341 tcatcaatga ttcattcact actgggtact gccatccaaa caatgatttt tatagttttc
     2401 ctaaaatctc cgttaaccca cacgcgttca ctgaaggcgg tcctgcgcaa tttgtgaatg
     2461 cgacgagcaa agaagtggtt gaatgggcgg ctaagttagg gcttccactc gtgtttagat
     2521 gggacgactc aaacgctcaa agaaaagaat acgccggttt gtaccacgaa gttgctcagg
     2581 cacatggtgt cgatgttagt caggttcgac acaagctgac gctgctggtc aaccaaaatg
     2641 tagatggtga agcagcaagg gcagaagctc gcgtgtattt ggaagagttt gtccgtgaat
     2701 cttactcaaa taccgacttt gagcaaaaaa tgggagagct gttgtcagaa aatgccatcg
     2761 gtacttatga agaaagtact caggcagcgc gagttgcgat tgagtgttgt ggtgccgcgg
     2821 acctattgat gtcttttgag tcgatggaag ataaagcgca gcaaagagcg gttatcgatg
     2881 tggtaaacgc caacatcgtc aaataccact cgtaaactag tgcatgcgtg acacaagaat
     2941 ccctgttact tctcgaccgt attgattcgg atgattccta cgcgagcctg cggaacgacc
     3001 aggaattctg ggagccgctg gcccgccgag ccctggagga gctcgggctg ccggtgccgc
     3061 cggtgctgcg ggtgcccggc gagagcacca accccgtact ggtcggcgag cccggcccgg
     3121 tgatcaagct gttcggcgag cactggtgcg gtccggagag cctcgcgtcg gagtcggagg
     3181 cgtacgcggt cctggcggac gccccggtgc cggtgccccg cctcctcggc cgcggcgagc
     3241 tgcggcccgg caccggagcc tggccgtggc cctacctggt gatgagccgg atgaccggca
     3301 ccacctggcg gtccgcgatg gacggcacga ccgaccggaa cgcgctgctc gccctggccc
     3361 gcgaactcgg ccgggtgctc ggccggctgc acagggtgcc gctgaccggg aacaccgtgc
     3421 tcacccccca ttccgaggtc ttcccggaac tgctgcggga acgccgcgcg gcgaccgtcg
     3481 aggaccaccg cgggtggggc tacctctcgc cccggctgct ggaccgcctg gaggactggc
     3541 tgccggacgt ggacacgctg ctggccggcc gcgaaccccg gttcgtccac ggcgacctgc
     3601 acgggaccaa catcttcgtg gacctggccg cgaccgaggt caccgggatc gtcgacttca
     3661 ccgacgtcta tgcgggagac tcccgctaca gcctggtgca actgcatctc aacgccttcc
     3721 ggggcgaccg cgagatcctg gccgcgctgc tcgacggggc gcagtggaag cggaccgagg
     3781 acttcgcccg cgaactgctc gccttcacct tcctgcacga cttcgaggtg ttcgaggaga
     3841 ccccgctgga tctctccggc ttcaccgatc cggaggaact ggcgcagttc ctctgggggc
     3901 cgccagacac cgcccccggc gcctaaaagc ttggcactgg ccgtcgtttt acaacgtcgt
     3961 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     4021 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4081 aatggcgaat ggcgataagc tagcccaacc agcccgcgaa cggctggcag ccagcgtaag
     4141 gcgcggctca tcgggcggcg ttcgccacga tgtcctgcac ttcgagccaa gcctcgaaca
     4201 cctgctggtg tgcacgactc acccggttgt tgacaccgcg cgcggccgtg cgggctcggt
     4261 ggggcggctc tgtcgccctt gccagcgtga gtagcgcgta cctcacctcg cccaacaggt
     4321 cgcacacagc cgattcgtac gccataaagc caggtgagcc caccagctcc gtaagttcgg
     4381 gcgctgtgtg gctcgtaccc gcgcattcag gcggcagggg gtctaacggg tctaaggcgg
     4441 cgtgtacggc cgccacagcg gctctcagcg gcccggaaac gtccttgaaa cgacgtatgt
     4501 gttcctcctg gttggtacag gtggttgggg gtgctcggct gtcgctggtg ttccaccacc
     4561 agggctcgac gggagagcgg gggagtgtgc agttgtgggg tggcccctca gcgaaatatc
     4621 tgacttggag ctcgtgtcgg accatacacc ggtgattaat cgtggtctac taccaagcgt
     4681 gagccacgtc gccgacgaat ttgagcagct ctggctgccg tactggccgc tggcaagcga
     4741 cgatctgctc gaggggatct accgccaaag ccgcgcgtcg gccctaggcc gccggtacat
     4801 cgaggcgaac ccaacagcgc tggcaaacct gctggtcgtg gacgtagacc atccagacgc
     4861 agcgctccga gcgctcagcg cccgggggtc ccatccgctg cccaacgcga tcgtgggcaa
     4921 tcgcgccaac ggccacgcac acgcagtgtg ggcactcaac gcccctgttc cacgcaccga
     4981 atacgcgcgg cgtaagccgc tcgcatacat ggcggcgtgc gccgaaggcc ttcggcgcgc
     5041 cgtcgacggc gaccgcagtt actcaggcct catgaccaaa aaccccggcc acatcgcctg
     5101 ggaaacggaa tggctccact cagatctcta cacactcagc cacatcgagg ccgagctcgg
     5161 cgcgaacatg ccaccgccgc gctggcgcca gcagaccacg tacaaagcgg ctccgacgcc
     5221 gctagggcgg aattgcgcac tgttcgattc cgtcaggttg tgggcctatc gtcccgccct
     5281 catgcggatc tacctgccga cccggaacgt ggacggactc ggccgcgcga tctatgccga
     5341 gtgccacgtc cgaaacgccg agttcccgtg caacgacgtg tgtcccggac cgctaccgga
     5401 cagcgaggtc cgcgccatcg ccaacagcat ttggcgttgg atcacaacca agtcgcgcat
     5461 ttgggcggac gggatcgtgg tctacgaggc cacactcagt gcgcgccagt cggccatctc
     5521 gcggaagggc gcagcagcgc gcacggcggc gagcacagtt gcgcggcgcg caaagtccgc
     5581 gtcagccatg gaggcattgc tatgagcgac ggctacagcg acggctacaa ccggcagccg
     5641 actgtccgca aaaagcggcg cgtgaccgcc gccgaaggcg ctcgaatcac cggactatcc
     5701 gaacgccacg tcgtccggct cgtggcgcag gaacgcagcg agtggctcgc cgagcaggct
     5761 gcacgccgcg aacgcatccg cgcctatcac gacgacgagg gccactcttg gccgcaaacg
     5821 gccaaacatt tcgggctgca tctggacacc gttaagcgac tcggctatcg ggcgaggaaa
     5881 gagcgtgcgg cagaacagga agcggctcaa aaggcccaca acgaagccga caatccaccg
     5941 ctgttctaac gcaattgggg agcgggtgtc gcgggggttc cgtggggggt tccgttgcaa
     6001 cgggtcggac aggtaaaagt cctggtagac gctagttttc tggtttgggc catgcctgtc
     6061 tcgttgcgtg tttcgttgcg cccgttttga ataccagcca gacgagacgg ggttctacga
     6121 atcttggtcg ataccaagcc atttccgctg aatatcgggg agctcaccgc cagaatcggt
     6181 ggttgtggtg atgtacgtgg cgaactccgt tgtagtgcct gtggtggcat ccgtggccac
     6241 tctcgttgca cggttcgttg tgccgttaca ggccccgttg acagctcacc gaacgtagtt
     6301 aaaacatgct ggtcaaacta ggtttaccaa cgatacgagt cagctcatct agggccagtt
     6361 ctaggcgttg ttcgttgcgc ggttcgttgc gcatgtttcg tgtggttgct agcttcacgc
     6421 tgccgcaagc actcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt
     6481 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa
     6541 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac
     6601 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
     6661 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
     6721 agggatcaag atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     6781 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6841 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6901 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actccaagac gaggcagcgc
     6961 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     7021 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     7081 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     7141 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     7201 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     7261 cgccagccga actgttcgcc aggctcaagg cgcggatgcc cgacggcgag gatctcgtcg
     7321 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     7381 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     7441 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     7501 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     7561 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     7621 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     7681 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaaaaggat
     7741 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7801 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7861 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7921 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7981 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     8041 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     8101 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     8161 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     8221 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     8281 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     8341 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     8401 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     8461 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     8521 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     8581 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     8641 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     8701 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     8761 ctttatgctt ccggctcgta tgttgtgtgg aattgtga
//
