LOCUS       sequence-11545-e        9385 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 9385)
  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 9385)
  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-Int
FEATURES             Location/Qualifiers
     source          1..9385
                     /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"
     terminator      4675..4721
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             7352..8146
                     /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(7406..7425)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8016..8035
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      8430..9018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8919..8938
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9172..9189
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9306..9327
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9342..9347,9348..9365,9366..9372)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(9380..9385,1..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(9385,1..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 ggtgtacgat cgactgccag gcatcaaata
     4681 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tgccatcatg gccgcggtga
     4741 tcagctagag ccgtgaacga cagggcgaac gccagcccgc cgacggcgag ggttccgacc
     4801 gctgcaactc ccggtgcaac cttgtcccgg tctattctct tcactgcacc agctccaatc
     4861 tggtgtgaat gcccctcgtc tgttcgcgca ggcggggggc tctattcgtt tgtcagcatc
     4921 gaaagtagcc agatcaggga tgcgttgcaa ccgcgtatgc ccaggtcaga agagtcgcac
     4981 aagagttgca gacccctgga aagaaaaatg gccagagggc gaaaacaccc tctgaccagc
     5041 ggagcgggcg acgggaatcg aacccgcgta gctagtttgg aagaatgggt gtctgccgac
     5101 cacatatatg ggccggtcaa gataggtttt taccccctct cggctgcatc ctctaagtgg
     5161 aaagaaattg caggtcgtag aagcgcgttg aagcctgaga gttgcacagg agttgcaacc
     5221 cggtagcctt gttcacgacg agaggagacc tagttggcac gtcgcggatg gggatcgctg
     5281 aagactcagc gcagcgggag gatccaagcc tcatacgtca acccgcagga cggtgtgagg
     5341 tactacgcgc tgcagaccta cgacaacaag atggacgccg aagcctggct cgcgggcgag
     5401 aagcggctca tcgagatgga gacctggacc cctccacagg accgggcgaa gaaggcagcc
     5461 gccagcgcca tcacgctgga ggagtacacc cggaagtggc tcgtggagcg cgacctcgca
     5521 gacggcacca gggatctgta cagcgggcac gcggagcgcc gcatctaccc ggtgctaggt
     5581 gaagtggcgg tcacagagat gacgccagct ctggtgcgtg cgtggtgggc cgggatgggt
     5641 aggaagcacc cgactgcccg ccggcatgcc tacaacgtcc tccgggcggt gatgaacaca
     5701 gcggtcgagg acaagctgat cgcagagaac ccgtgccgga tcgagcagaa ggcagccgat
     5761 gagcgcgacg tagaggcgct gacgcctgag gagctggaca tcgtcgccgc tgagatcttc
     5821 gagcactacc ggatcgcggc atacatcctg gcgtggacga gcctccggtt cggagagctg
     5881 atcgagcttc gccgcaagga catcgtggac gacggcatga cgatgaagct ccgggtgcgc
     5941 cgtggcgctt cccgcgtggg gaacaagatc gtcgttggca acgccaagac cgtccggtcg
     6001 aagcgtcctg tgacggttcc gcctcacgtc gcggagatga tccgagcgca catgaaggac
     6061 cgtacgaaga tgaacaaggg ccccgaggca ttcctggtga ccacgacgca gggcaaccgg
     6121 ctgtcgaagt ccgcgttcac caagtcgctg aagcgtggct acgccaagat cggtcggccg
     6181 gaactccgca tccacgacct ccgcgctgtc ggcgctacgt tcgccgctca ggcaggtgcg
     6241 acgaccaagg agctgatggc ccgtctcggt cacacgactc ctaggatggc gatgaagtac
     6301 cagatggcgt ctgaggcccg cgacgaggct atcgctgagg cgatgtccaa gctggccaag
     6361 acctcctgaa acgcaaaaag cccccctccc aaggacactg agtcctaaag aggggggttt
     6421 cttgtcagta cgcgaagaac cacgcctggc cgcgagcgcc agcaccgccg ctctgtgcgg
     6481 agacctgggc accagccccg ccgccgccag gagcattgcc gttcccgcca gctagcaaca
     6541 attggggagc gggtgtcgcg ggggttccgt ggggggttcc gttgcaacgg gtcggacagg
     6601 taaaagtcct ggtagacgct agttttctgg tttgggccat gcctgtctcg ttgcgtgttt
     6661 cgttgcgccc gttttgaata ccagccagac gagacggggt tctacgaatc ttggtcgata
     6721 ccaagccatt tccgctgaat atcggggagc tcaccgccag aatcggtggt tgtggtgatg
     6781 tacgtggcga actccgttgt agtgcctgtg gtggcatccg tggccactct cgttgcacgg
     6841 ttcgttgtgc cgttacaggc cccgttgaca gctcaccgaa cgtagttaaa acatgctggt
     6901 caaactaggt ttaccaacga tacgagtcag ctcatctagg gccagttcta ggcgttgttc
     6961 gttgcgcggt tcgttgcgca tgtttcgtgt ggttgctagc ttcacgctgc cgcaagcact
     7021 cagggcgcaa gggctgctaa aggaagcgga acacgtagaa agccagtccg cagaaacggt
     7081 gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa
     7141 agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat
     7201 ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct
     7261 gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg gatcaagatc
     7321 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     7381 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     7441 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     7501 gaccgacctg tccggtgccc tgaatgaact ccaagacgag gcagcgcggc tatcgtggct
     7561 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     7621 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     7681 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     7741 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     7801 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     7861 gttcgccagg ctcaaggcgc ggatgcccga cggcgaggat ctcgtcgtga cccatggcga
     7921 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     7981 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     8041 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     8101 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     8161 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     8221 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     8281 cagcgcgggg atctcatgct ggagttcttc gcccacccca aaaggatcta ggtgaagatc
     8341 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8401 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     8461 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     8521 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     8581 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     8641 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     8701 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     8761 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     8821 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     8881 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     8941 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     9001 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     9061 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     9121 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     9181 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     9241 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     9301 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     9361 gctcgtatgt tgtgtggaat tgtga
//
