LOCUS       sequence-207440-        5698 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5698)
  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 5698)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pLINK cyclin T1 (1-280) myc (all 200) (P#575)
FEATURES             Location/Qualifiers
     source          1..5698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      26..614
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     515..534
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     768..785
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    902..923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(938..943,944..961,962..968)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    976..992
                     /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     981..1003
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1000..1016
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1000..1016
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1130..2308
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1361..2299
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2256..2276
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3241..3270
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3393..3787
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(4098..4115)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4098..4114)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4107..4129)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4323..4342)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4442..4464
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4502..4520)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4588..4692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4693..4761,4762..5553)
                     /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(4911..4930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
       61 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      121 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      181 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      241 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      301 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      361 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      421 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      481 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      541 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      601 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      661 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      721 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      781 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
      841 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
      901 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
      961 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga
     1021 ttacgaatta attcgagctc gcccagcttt gcaaagatgg ataaagtttt aaacagagag
     1081 gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc
     1141 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     1201 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     1261 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     1321 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
     1381 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
     1441 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
     1501 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
     1561 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
     1621 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
     1681 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
     1741 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
     1801 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
     1861 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     1921 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     1981 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     2041 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     2101 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     2161 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     2221 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     2281 tttttttctt ccatttcagg tgtcgtgagg aattagcttg cttacatttg cttctgacac
     2341 aactgtgttc actagcaacc tcaaacagac accatggccg gatccaccat ggagggagag
     2401 aggaagaaca acaacaaacg gtggtatttc actcgagaac agctggaaaa tagcccatcc
     2461 cgtcgttttg gcgtggaccc agataaagaa ctttcttatc gccagcaggc ggccaatctg
     2521 cttcaggaca tggggcagcg tcttaacgtc tcacaattga ctatcaacac tgctatagta
     2581 tacatgcatc gattctacat gattcagtcc ttcacacagt tccctggaaa ttctgtggct
     2641 ccagcagcct tgtttctagc agctaaagtg gaggagcagc ccaaaaaatt ggaacatgtc
     2701 atcaaggtag cacatacttg tctccatcct caggaatccc ttcctgatac tagaagtgag
     2761 gcttatttgc aacaagttca agatctggtc attttagaaa gcataatttt gcagacttta
     2821 ggctttgaac taacaattga tcacccacat actcatgtag taaagtgcac tcaacttgtt
     2881 cgagcaagca aggacttagc acagacttct tacttcatgg caaccaacag cctgcatttg
     2941 accacattta gcctgcagta cacacctcct gtggtggccg ctgtcgctat tgcgctagct
     3001 gccaagtggt ccaattggga gatcccagtc tcaactgacg ggaagcactg gtgggagtat
     3061 gttgacgcca ctgtgacctt ggaactttta gatgaactga cacatgagtt tctacagatt
     3121 ttggagaaaa ctcccaacag gctcaaacgc atttggaatt ggagggcatg cgaggctgcc
     3181 aagaaaacaa aagcagatga ccgaggaaca gatgaaaaga cttcagagga tatcatgccg
     3241 gagcagaagc tgatatccga ggaggacctg gccatgtaag aattcctcga gatcgattag
     3301 actagtgcta gcaattcacc ccaccagtgc aggctgccta tcagaaagtg gtggctggtg
     3361 tggctaatgc cctggcccac aagtatcact aagctcgctt tcttgctgtc caatttctat
     3421 taaaggttcc tttgttccct aagtccaact actaaactgg gggatattat gaagggcctt
     3481 gagcatctgg attctgccta ataaaaaaca tttattttca ttgcaatgat gtatttaaat
     3541 tatttctgaa tattttacta aaaagggaat gtgggaggtc agtgcattta aaacataaag
     3601 aaatgaagag ctagttcaaa ccttgggaaa atacactata tcttaaactc catgaaagaa
     3661 ggtgaggctg caaacagcta atgcacattg gcaacagccc ctgatgccta tgccttattc
     3721 atccctcaga aaaggattca agtagaggct tgatttggag gttaaagttt tgctatgctg
     3781 tattttacat tacttattgt tttagctgtc ctcatgaatg tcttttcact acccatttgc
     3841 ttatcctgca tctctcagcc ttgactccac tcagttctct tgcttagaga taccaccttt
     3901 cccctgaagt gttccttcca tgttttacgg cgagatggtt tctcctcgcc tggccactca
     3961 gccttagttg tctctgttgt cttatagagg tctacttgaa gaaggaaaaa cagggggcat
     4021 ggtttgactg tcctgtgagc ccttcttccc tgcctccccc actcacagtg acccggaatc
     4081 tgcagcccaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     4141 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4201 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     4261 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     4321 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4381 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4441 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     4501 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4561 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4621 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4681 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4741 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4801 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4861 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4921 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4981 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5041 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5101 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5161 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5221 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5281 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5341 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5401 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5461 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5521 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5581 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5641 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacc
//
