LOCUS       sequence-436921-        4710 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of H. pylori J99 methylthioadenosine nucleosidase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4710)
  AUTHORS   Wang S, Haapalainen AM, Yan F, Du Q, Tyler PC, Evans GB, 
            Rinaldo-Matthis A, Brown RL, Norris GE, Almo SC, Schramm VL
  TITLE     A picomolar transition state analogue inhibitor of MTAN as a 
            specific antibiotic for Helicobacter pylori.
  JOURNAL   Biochemistry. 2012 Sep 4;51(35):6892-4. doi: 10.1021/bi3009664. Epub
            2012 Aug 22.
  PUBMED    22891633
REFERENCE   2  (bases 1 to 4710)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/bi3009664";
            journalName: "Biochemistry"; date: "2012-09-4- 4"; volume: "51"; 
            issue: "35"; pages: "6892-4"
COMMENT     Sequence Label: Hp MTAN
FEATURES             Location/Qualifiers
     source          1..4710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     858..875
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     859..875
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1338..1357
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1338..1356
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1357..1381
                     /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)."
     CDS             1410..1427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1428..1448
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2152..2199
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2422..2441)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2447..2466)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        2508..2599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2600..2668,2669..3460)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2818..2837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3543..4625
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3562..3581)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactcct tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag cccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag gcgagagtag
      781 ggaactgcca ggcatcaaac taagcagaag gcccctgacg gatggccttt ttgcgtttct
      841 acaaactctt tctgtgttgt aaaacgacgg ccagtcttaa gctcgggccc cctgggcggt
      901 tctgataacg agtaatcgtt aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt
      961 ttatgggggg agtttaggga aagagcattt gtcagaatat ttaagggcgc ctgtcacttt
     1021 gcttgatata tgagaattat ttaaccttat aaatgagaaa aaagcaacgc actttaaata
     1081 agatacgttg ctttttcgat tgatgaacac ctataattaa actattcatc tattatttat
     1141 gattttttgt atatacaata tttctagttt gttaaagaga attaagaaaa taaatctcga
     1201 aaataataaa gggaaaatca gtttttgata tcaaaattat acatgtcaac gataatacaa
     1261 aatataatac aaactataag atgttatcag tatttattat catttagaat aaattttgtg
     1321 tcgcccttcc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     1381 ccctctagaa ggaggtaaaa catatgggcc atcaccatca ccatcacgaa aacctgtact
     1441 tccaaggtgt gcaaaaaatt ggcattttag gggcgatgag agaagaaata acccctatac
     1501 tagaattgtt tggcgtggat tttgaagaga tccctttagg ggggaatgtt ttccataaag
     1561 gcgtttatca taataaggaa atcattgtcg cttatagcaa gattggcaag gtgcattcca
     1621 ctttaaccac aacaagcatg attttagcgt ttggcgttca gaaggtgctt tttagcgggg
     1681 tggctggaag cttagttaaa gatttaaaaa tcaatgattt gttagtggct actcaattag
     1741 tccagcacga tgtggatttg agcgcgtttg atcacccttt agggtttatc cccgaaagcg
     1801 cgatttttat tgaaacgagt ggaagtttaa acgctttagc taaaaagatc gctaatgagc
     1861 aacatatcgc gctcaaagaa ggcgtcatcg catcaggcga tcagtttgtg catagcaaag
     1921 aaaggaaaga atttttagtt agcgagttta aagcgagcgc ggtggaaatg gagggggcga
     1981 gcgtggcgtt tgtgtgccaa aaatttggcg tgccatgctg cgtgctaagg agcattagcg
     2041 ataacgccga tgaaaaagcc ggtatgagtt ttgatgaatt tttagaaaaa agcgctcaca
     2101 cttcagcgaa attcttaaaa agcatggtgg atgagcttta actcgagccc cctagcataa
     2161 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc ccctgagacg cgtcaatcga
     2221 gttcgtacct aagggcgaca ccccataatt agcccgggcg aaaggcccag tctttcgact
     2281 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcatgggg agtccccaca
     2341 ctaccatcgg cgctacggcg tttcacttct gagttcggca tggggtcagg tgggaccacc
     2401 gcgctactgc cgccaggcaa acaaggggtg ttatgagcca tattcaggta taaatgggct
     2461 cgcgataatg ttcagaattg gttaattggt tgtaacactg acccctattt gtttattttt
     2521 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2581 atattgaaaa aggaagaata tgagtattca acatttccgt gtcgccctta ttcccttttt
     2641 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2701 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2761 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2821 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2881 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2941 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3001 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3061 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3121 cgagcgtgac accacgatgc ctgtagcgat ggcaacaacg ttgcgcaaac tattaactgg
     3181 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3241 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatccgg
     3301 agccggtgag cgtggttctc gcggtatcat cgcagcgctg gggccagatg gtaagccctc
     3361 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3421 gatcgctgag ataggtgcct cactgattaa gcattggtaa gcggcgcgcc atcgaatggc
     3481 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     3541 atatgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     3601 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     3661 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     3721 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     3781 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
     3841 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
     3901 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     3961 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     4021 ttttctccca tgaggacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     4081 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     4141 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
     4201 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
     4261 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
     4321 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagatagct
     4381 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     4441 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     4501 cagtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
     4561 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4621 agtgactcat gaccaaaatc ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc
     4681 agaccccgta gaaaagatca aaggatcttc
//
