LOCUS       sequence-142533-        7476 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 7476)
  AUTHORS   Taube R, Lin X, Irwin D, Fujinaga K, Peterlin BM
  TITLE     Interaction between P-TEFb and the C-terminal domain of RNA 
            polymerase II activates transcriptional elongation from sites 
            upstream or downstream of target genes.
  JOURNAL   Mol Cell Biol. 2002 Jan . 22(1):321-31.
  PUBMED    11739744
REFERENCE   2  (bases 1 to 7476)
  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: "Mol Cell 
            Biol. 2002 Jan . 22(1):321-31."
COMMENT     Sequence Label: pEF myc Gal cyclin T1 (1-726) (P#1026)
FEATURES             Location/Qualifiers
     source          1..7476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2336..3514
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2567..3505
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3462..3482
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3586..3603
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3613..3642
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3658..4098
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     polyA_signal    6377..6771
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(7082..7099)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7082..7098)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7091..7113)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7307..7326)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7426..7448
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 acatgattac gaattaattc gagctcgccc agctttgcaa agatggataa agttttaaac
     2281 agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc
     2341 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     2401 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     2461 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     2521 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg
     2581 tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc
     2641 cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
     2701 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
     2761 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
     2821 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
     2881 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
     2941 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
     3001 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
     3061 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     3121 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     3181 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     3241 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     3301 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     3361 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     3421 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     3481 tcaaagtttt tttcttccat ttcaggtgtc gtgaggaatt agcttgctta catttgcttc
     3541 tgacacaact gtgttcacta gcaacctcaa acagacacca tgccgcatca ccaccatcat
     3601 cacgccatgc cggagcagaa gctgatatcc gaggaggacc tggccatggc cggatccatg
     3661 aagctactgt cttctatcga acaagcatgc gatatttgcc gacttaaaaa gctcaagtgc
     3721 tccaaagaaa aaccgaagtg cgccaagtgt ctgaagaaca actgggagtg tcgctactct
     3781 cccaaaacca aaaggtctcc gctgactagg gcacatctga cagaagtgga atcaaggcta
     3841 gaaagactgg aacagctatt tctactgatt tttcctcgag aagaccttga catgattttg
     3901 aaaatggatt ctttacagga tataaaagca ttgttaacag gattatttgt acaagataat
     3961 gtgaataaag atgccgtcac agatagattg gcttcagtgg agactgatat gcctctaaca
     4021 ttgagacagc atagaataag tgcgacatca tcatcggaag agagtagtaa caaaggtcaa
     4081 agacagttga ctgtatcgcc ggaattcccc atggagggag agaggaagaa caacaacaaa
     4141 cggtggtatt tcactcgaga acagctggaa aatagcccat cccgtcgttt tggcgtggac
     4201 ccagataaag aactttctta tcgccagcag gcggccaatc tgcttcagga catggggcag
     4261 cgtcttaacg tctcacaatt gactatcaac actgctatag tatacatgca tcgattctac
     4321 atgattcagt ccttcacaca gttccctgga aattctgtgg ctccagcagc cttgtttcta
     4381 gcagctaaag tggaggagca gcccaaaaaa ttggaacatg tcatcaaggt agcacatact
     4441 tgtctccatc ctcaggaatc ccttcctgat actagaagtg aggcttattt gcaacaagtt
     4501 caagatctgg tcattttaga aagcataatt ttgcagactt taggctttga actaacaatt
     4561 gatcacccac atactcatgt agtaaagtgc actcaacttg ttcgagcaag caaggactta
     4621 gcacagactt cttacttcat ggcaaccaac agcctgcatt tgaccacatt tagcctgcag
     4681 tacacacctc ctgtggtggc ctgtgtctgc attcacctgg cttgcaagtg gtccaattgg
     4741 gagatcccag tctcaactga cgggaagcac tggtgggagt atgttgacgc cactgtgacc
     4801 ttggaacttt tagatgaact gacacatgag tttctacaga ttttggagaa aactcccaac
     4861 aggctcaaac gcatttggaa ttggagggca tgcgaggctg ccaagaaaac aaaagcagat
     4921 gaccgaggaa cagatgaaaa gacttcagag cagacaatcc tcaatatgat ttcccagagc
     4981 tcttcagaca caaccattgc aggtttaatg agcatgtcaa cttctaccac aagtgcagtg
     5041 ccttccctgc cagtctccga agagtcatcc agcaacttaa ccagtgtgga gatgttgccg
     5101 ggcaagcgtt ggctgtcctc ccaaccttct ttcaaactag aacctactca gggtcatcgg
     5161 actagtgaga atttagcact tacaggagtt gatcattcct taccacagga tggttcaaat
     5221 gcatttattt cccagaagca gaatagtaag agtgtgccat cagctaaagt gtcactgaaa
     5281 gaataccgcg cgaagcatgc agaagaattg gctgcccaga agaggcaact ggagaacatg
     5341 gaagccaatg tgaagtcaca atatgcatat gctgcccaga atctcctttc tcatcatgat
     5401 agccattctt cagtcattct aaaaatgccc atagagggtt cagaaaaccc cgagcggcct
     5461 tttctggaaa aggctgacaa aacagctctc aaaatgagaa tcccagtggc aggtggagat
     5521 aaagctgcgt cttcaaaacc agaggagata aaaatgcgca taaaagtcca tgctgcagct
     5581 gataagcaca attctgtaga ggacagtgtt acaaagagcc gagagcacaa agaaaagcac
     5641 aagactcacc catctaatca tcatcgtcat cataatcacc actcacacaa gcactctcat
     5701 tcccaacttc cagttggtac tgggaacaaa cgtcctggtg atccaaaaca tagtagccag
     5761 acaagcaact tagcacataa aacctatagc ttgtctagtt cttttccctc ttccagttct
     5821 actcgtaaaa ggggaccctc tgaagagact ggaggggctg tgtttgatca tccagccaag
     5881 attgccaaga gtactaaatc ctcttcccta aatttctcct tcccttcact tcctacaatg
     5941 ggtcagatgc ctgggcatag ctcagacaca agtggccttt ccttttcaca gcccagctgt
     6001 aaaactcgtg tccctcattc gaaactggat aaagggccca ctggggccaa tggtcacaac
     6061 acgacccaga caatagacta tcaagacact gtgaatatgc ttcactccct gctcagtgcc
     6121 cagggtgttc agcccactca gcctactgca tttgaatttg ttcgtcctta tagtgactat
     6181 ctgaatcctc ggtctggtgg aatctcctcg agatctggca atacagacaa accccggcca
     6241 ccatctctgc catcagaacc tcctccacca cttccaccct tcctaagtaa tctagaaatt
     6301 caccccacca gtgcaggctg cctatcagaa agtggtggct ggtgtggcta atgccctggc
     6361 ccacaagtat cactaagctc gctttcttgc tgtccaattt ctattaaagg ttcctttgtt
     6421 ccctaagtcc aactactaaa ctgggggata ttatgaaggg ccttgagcat ctggattctg
     6481 cctaataaaa aacatttatt ttcattgcaa tgatgtattt aaattatttc tgaatatttt
     6541 actaaaaagg gaatgtggga ggtcagtgca tttaaaacat aaagaaatga agagctagtt
     6601 caaaccttgg gaaaatacac tatatcttaa actccatgaa agaaggtgag gctgcaaaca
     6661 gctaatgcac attggcaaca gcccctgatg cctatgcctt attcatccct cagaaaagga
     6721 ttcaagtaga ggcttgattt ggaggttaaa gttttgctat gctgtatttt acattactta
     6781 ttgttttagc tgtcctcatg aatgtctttt cactacccat ttgcttatcc tgcatctctc
     6841 agccttgact ccactcagtt ctcttgctta gagataccac ctttcccctg aagtgttcct
     6901 tccatgtttt acggcgagat ggtttctcct cgcctggcca ctcagcctta gttgtctctg
     6961 ttgtcttata gaggtctact tgaagaagga aaaacagggg gcatggtttg actgtcctgt
     7021 gagcccttct tccctgcctc ccccactcac agtgacccgg aatctgcagc ccaagcttgg
     7081 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     7141 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     7201 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     7261 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     7321 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     7381 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     7441 gtcagaggtt ttcaccgtca tcaccgaaac gcgcga
//
