LOCUS       sequence-464335-        4886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human 5'-methylthioadenosine phosphorylase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4886)
  AUTHORS   Guan R, Ho MC, Brenowitz M, Tyler PC, Evans GB, Almo SC, Schramm VL
  TITLE     Entropy-driven binding of picomolar transition state analogue 
            inhibitors to human 5'-methylthioadenosine phosphorylase.
  JOURNAL   Biochemistry. 2011 Nov 29;50(47):10408-17. doi: 10.1021/bi201321x. 
            Epub 2011 Nov 7.
  PUBMED    21985704
REFERENCE   2  (bases 1 to 4886)
  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: "10.1021/bi201321x";
            journalName: "Biochemistry"; date: "2011-11-29- 29"; volume: "50"; 
            issue: "47"; pages: "10408-17"
COMMENT     Sequence Label: human MTAP
FEATURES             Location/Qualifiers
     source          1..4886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..78
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        59..77
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    78..102
                     /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             147..164
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             165..185
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      1049..1096
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1319..1338)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1344..1363)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        1405..1496
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1497..1565,1566..2357)
                     /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(1715..1734)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             2440..3522
                     /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(2459..2478)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3608..4196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4097..4116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4350..4367
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4465..4482
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4466..4482
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gatgttatca gtatttatta tcatttagaa taaattttgt gtcgcccttc cgcgaaatta
       61 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg
      121 tttaactttt aggaggataa catatgcatc atcaccatca ccacgagaac ctgtacttcc
      181 aaggcatggc gagcggtacc accaccaccg ccgtcaaaat cggtatcatt ggcggtacgg
      241 gtctggatga cccggaaatc ctggagggtc gcacggagaa gtacgttgac acgccattcg
      301 gcaagccgtc tgatgcgctg attctgggca agattaagaa tgtggattgc gtgctgctgg
      361 ctcgtcatgg tcgtcagcat acgattatgc cgagcaaagt gaactatcag gcgaacattt
      421 gggcactgaa agaagagggc tgtacgcacg tgatcgttac gaccgcgtgc ggcagcctgc
      481 gcgaagagat tcaaccaggt gatatcgtca tcattgacca gttcattgat cgtaccacga
      541 tgcgtccgca atccttttac gacggttccc atagctgtgc gcgtggcgtc tgccacatcc
      601 cgatggcaga accgttttgt ccgaaaaccc gtgaggttct gatcgaaacc gctaagaagc
      661 tgggcttgcg ctgccacagc aagggtacga tggtcaccat cgagggtcct cgctttagca
      721 gccgtgcgga gagctttatg tttcgcacct ggggtgcaga cgttatcaat atgaccacgg
      781 ttccggaagt tgtgctggca aaagaagccg gtatctgcta tgccagcatt gcgatggcga
      841 ccgactacga ttgttggaaa gagcacgaag aagcagtgag cgtcgaccgt gttctgaaaa
      901 ctctgaaaga gaatgccaat aaggctaaat ctctgctgtt gaccacgatt ccgcagattg
      961 gttccactga gtggtcggaa accctgcaca acctgaaaaa catggcacag ttctctgtgc
     1021 tgttgccgcg tcactgataa gaattccccc agcataaccc cttggggcct ctaaacgggt
     1081 cttgaggggt tttttgcccc tgagacgcgt caatcgagtt cgtacctaag ggcgacaccc
     1141 cataattagc ccgggcgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc
     1201 ctggcagttc cctactctcg catggggagt ccccacacta ccatcggcgc tacggcgttt
     1261 cacttctgag ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaaca
     1321 aggggtgtta tgagccatat tcaggtataa atgggctcgc gataatgttc agaattggtt
     1381 aattggttgt aacactgacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1441 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagaatatga
     1501 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1561 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1621 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1681 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     1741 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1801 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1861 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1921 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1981 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2041 tagcgatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2101 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2161 cccttccggc tggctggttt attgctgata aatccggagc cggtgagcgt ggttctcgcg
     2221 gtatcatcgc agcgctgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2281 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2341 tgattaagca ttggtaagcg gcgcgccatc gaatggcgca aaacctttcg cggtatggca
     2401 tgatagcgcc cggaagagag tcaattcagg gtggtgaata tgaaaccagt aacgttatac
     2461 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     2521 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     2581 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     2641 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     2701 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     2761 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     2821 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     2881 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga ggacggtacg
     2941 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     3001 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
     3061 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
     3121 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
     3181 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
     3241 atctcggtag tgggatacga cgataccgaa gatagctcat gttatatccc gccgttaacc
     3301 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
     3361 tctcagggcc aggcggtgaa gggcaatcag ctgttgccag tctcactggt gaaaagaaaa
     3421 accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3481 cagctggcac gacaggtttc ccgactggaa agcgggcagt gactcatgac caaaatccct
     3541 taacgtgagt tacgcgcgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3601 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3661 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3721 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttagccc
     3781 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3841 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3901 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3961 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4021 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4081 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4141 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4201 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4261 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4321 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaaggcg
     4381 agagtaggga actgccaggc atcaaactaa gcagaaggcc cctgacggat ggcctttttg
     4441 cgtttctaca aactctttct gtgttgtaaa acgacggcca gtcttaagct cgggccccct
     4501 gggcggttct gataacgagt aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg
     4561 ggttttttta tggggggagt ttagggaaag agcatttgtc agaatattta agggcgcctg
     4621 tcactttgct tgatatatga gaattattta accttataaa tgagaaaaaa gcaacgcact
     4681 ttaaataaga tacgttgctt tttcgattga tgaacaccta taattaaact attcatctat
     4741 tatttatgat tttttgtata tacaatattt ctagtttgtt aaagagaatt aagaaaataa
     4801 atctcgaaaa taataaaggg aaaatcagtt tttgatatca aaattataca tgtcaacgat
     4861 aatacaaaat ataatacaaa ctataa
//
