LOCUS       sequence-176944-        5724 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 5724)
  AUTHORS   Fujinaga K, Taube R, Wimmer J, Cujec TP, Peterlin BM
  TITLE     Interactions between human cyclin T, Tat, and the transactivation 
            response element (TAR) are disrupted by a cysteine to tyrosine 
            substitution found in mouse cyclin T.
  JOURNAL   Proc Natl Acad Sci U S A. 1999 Feb 16. 96(4):1285-90.
  PUBMED    9990016
REFERENCE   2  (bases 1 to 5724)
  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: "Proc Natl 
            Acad Sci U S A. 1999 Feb 16. 96(4):1285-90."
COMMENT     Sequence Label: pGEX-2TK cyclin T1 (1-250) (P#467)
FEATURES             Location/Qualifiers
     source          1..5724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(13..18,19..34,35..41)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    49..65
                     /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             88..741
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     699..721
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             748..765
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1604..1626)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1771..1789)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1857..1961
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1962..2030,2031..2822)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2180..2199)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2993..3581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3482..3501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3735..3752
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3825..3902
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3903..4985
                     /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(3922..3941)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5034..5039,5040..5057,5058..5064)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5072..5088
                     /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     5096..5112
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5096..5112
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5108..5635
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5128..5145)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5128..5144)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5137..5159)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tgaaatgagc tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat
       61 aacaatttca cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag
      121 ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat
      181 ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag
      241 tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc
      301 atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag
      361 atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat
      421 agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa
      481 atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct
      541 gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat
      601 gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag
      661 tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt
      721 ggtggcgacc atcctccaaa atcggatctg gttccgcgtg gatctcgtcg tgcatctgtt
      781 ggatccatgg agggagagag gaagaacaac aacaaacggt ggtatttcac tcgagaacag
      841 ctggaaaata gcccatcccg tcgttttggc gtggacccag ataaagaact ttcttatcgc
      901 cagcaggcgg ccaatctgct tcaggacatg gggcagcgtc ttaacgtctc acaattgact
      961 atcaacactg ctatagtata catgcatcga ttctacatga ttcagtcctt cacacagttc
     1021 cctggaaatt ctgtggctcc agcagccttg tttctagcag ctaaagtgga ggagcagccc
     1081 aaaaaattgg aacatgtcat caaggtagca catacttgtc tccatcctca ggaatccctt
     1141 cctgatacta gaagtgaggc ttatttgcaa caagttcaag atctggtcat tttagaaagc
     1201 ataattttgc agactttagg ctttgaacta acaattgatc acccacatac tcatgtagta
     1261 aagtgcactc aacttgttcg agcaagcaag gacttagcac agacttctta cttcatggca
     1321 accaacagcc tgcatttgac cacatttagc ctgcagtaca cacctcctgt ggtggcctgt
     1381 gtctgcattc acctggcttg caagtggtcc aattgggaga tcccagtctc aactgacggg
     1441 aagcactggt gggagtatgt tgacgccact gtgaccttgg aacttttaga tgaactgaca
     1501 catgagtttc tacagatttt ggagaaaact cccaacaggc tcaaagaatt catcgtgact
     1561 gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1621 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1681 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     1741 gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     1801 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     1861 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1921 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1981 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2041 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2101 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2161 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     2221 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2281 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2341 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2401 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2461 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     2521 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2581 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2641 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2701 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2761 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2821 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2881 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2941 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3121 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3601 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3661 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3721 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     3781 ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggc
     3841 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     3901 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     3961 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     4021 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     4081 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     4141 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
     4201 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
     4261 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     4321 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     4381 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     4441 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     4501 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
     4561 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
     4621 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
     4681 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct
     4741 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     4801 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     4861 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
     4921 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4981 agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca ggctttacac
     5041 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     5101 aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga
     5161 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     5221 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     5281 atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga
     5341 tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc
     5401 gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga
     5461 gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg
     5521 ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac ggtgcaccaa
     5581 tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac
     5641 tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
     5701 tcataacggt tctggcaaat attc
//
