LOCUS       sequence-13306-e        6609 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 6609)
  AUTHORS   Wang Y, Dow EC, Liang YY, Ramakrishnan R, Liu H, Sung TL, Lin X, 
            Rice AP
  TITLE     Phosphatase PPM1A regulates phosphorylation of Thr-186 in the Cdk9 
            T-loop.
  JOURNAL   J Biol Chem. 2008 Nov 28. 283(48):33578-84.
  PUBMED    18829461
REFERENCE   2  (bases 1 to 6609)
  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: "J Biol 
            Chem. 2008 Nov 28. 283(48):33578-84."
COMMENT     Sequence Label: HA-Cdk9 T186A
FEATURES             Location/Qualifiers
     source          1..6609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             898..978
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     promoter        complement(2161..2179)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2162..2179)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2199..2216)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2205..2429
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2475..2903
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2562..2581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2772..2793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2912..2932)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2917..3247
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3098..3233
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3160..3179
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3314..4108
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3368..3387)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3978..3997
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4282..4403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4319..4338)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4373..4392
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4452..4468)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4452..4468)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4465..4487)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4476..4492
                     /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)."
     promoter        complement(join(4500..4506,4507..4524,4525..4530))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4545..4566
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4683..4700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4854..5442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4934..4953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5613..6404,6405..6473))
                     /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     6236..6255
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6474..6578)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttatgtac
      901 ccatacgacg tcccagacta cgcttaccca tacgacgtcc cagactacgc ttacccatac
      961 gacgtcccag actacgctac cggtgcaaag cagtacgact cggtggagtg ccctttttgt
     1021 gatgaagttt ccaaatacga gaagctcgcc aagatcggcc aaggcacctt cggggaggtg
     1081 ttcaaggcca ggcaccgcaa gaccggccag aaggtggctc tgaagaaggt gctgatggaa
     1141 aacgagaagg aggggttccc cattacagcc ttgcgggaga tcaagatcct tcagcttcta
     1201 aaacacgaga atgtggtcaa cttgattgag atttgtcgaa ccaaagcttc cccctataac
     1261 cgctgcaagg gtagtatata cctggtgttc gacttctgcg agcatgacct tgctgggctg
     1321 ttgagcaatg ttttggtcaa gttcacgctg tctgagatca agagggtgat gcagatgctg
     1381 cttaacggcc tctactacat ccacagaaac aagatcctgc atagggacat gaaggctgct
     1441 aatgtgctta tcactcgtga tggggtcctg aagctggcag actttgggct ggcccgggcc
     1501 ttcagcctgg ccaagaacag ccagcccaac cgctacgcca accgtgtggt gacactctgg
     1561 taccggcccc cggagctgtt gctcggggag cgggactacg gcccccccat tgacctgtgg
     1621 ggtgctgggt gcatcatggc agagatgtgg acccgcagcc ccatcatgca gggcaacacg
     1681 gagcagcacc aactcgccct catcagtcag ctctgcggct ccatcacccc tgaggtgtgg
     1741 ccaaacgtgg acaactatga gctgtacgaa aagctggagc tggtcaaggg ccagaagcgg
     1801 aaggtgaagg acaggctgaa ggcctatgtg cgtgacccat acgcactgga cctcatcgac
     1861 aagctgctgg tgctggaccc tgcccagcgc atcgacagcg atgacgccct caaccacgac
     1921 ttcttctggt ccgaccccat gccctccgac ctcaagggca tgctctccac ccacctgacg
     1981 tccatgttcg agtacttggc accaccgcgc cggaagggca gccagatcac ccagcagtcc
     2041 accaaccaga gtcgcaatcc cgccaccacc aaccagacgg agtttgagcg cgtcttctga
     2101 gaattctgca gatatccatc acactggcgg ccgctcgagc atgcatctag agggccctat
     2161 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     2221 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2281 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2341 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2401 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2461 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2521 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2581 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     2641 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2701 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2761 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2821 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     2881 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     2941 tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3001 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3061 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3121 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3181 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3241 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     3301 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3361 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3421 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3481 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3541 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3601 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3661 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3721 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3781 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3841 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3901 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3961 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4021 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4081 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4141 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4201 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4261 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4321 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4381 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4441 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4501 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4561 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4621 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4681 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4741 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4801 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4861 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4921 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4981 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5041 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5101 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5161 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5221 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5281 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5341 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5401 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5461 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5521 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5581 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5641 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5701 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5761 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5821 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5881 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5941 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6001 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6061 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6121 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6181 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6241 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6301 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6361 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6421 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6481 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6541 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6601 cctgacgtc
//
