LOCUS       sequence-13304-e        5758 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 5758)
  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 5758)
  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: FLAG-Cdk9 T186A
FEATURES             Location/Qualifiers
     source          1..5758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             937..960
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2099..2721
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2298..2317)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2745..2765)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2750..3079
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2930..3065
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3117..3136)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3118..3136)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3150..3166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3150..3166)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3163..3185)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3174..3190
                     /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(3198..3204,3205..3222,3223..3228))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3243..3264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3381..3398)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3552..4140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3632..3651)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4311..5102,5103..5171))
                     /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     4934..4953
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5172..5276)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5303..5731
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5390..5409)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5600..5621
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tccaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attcagatct accatggact acaaagacga tgacgacaag
      961 cttgcggccg cgatggcaaa gcagtacgac tcggtggagt gccctttttg tgatgaagtt
     1021 tccaaatacg agaagctcgc caagatcggc caaggcacct tcggggaggt gttcaaggcc
     1081 aggcaccgca agaccggcca gaaggtggct ctgaagaagg tgctgatgga aaacgagaag
     1141 gaggggttcc ccattacagc cttgcgggag atcaagatcc ttcagcttct aaaacacgag
     1201 aatgtggtca acttgattga gatttgtcga accaaagctt ccccctataa ccgctgcaag
     1261 ggtagtatat acctggtgtt cgacttctgc gagcatgacc ttgctgggct gttgagcaat
     1321 gttttggtca agttcacgct gtctgagatc aagagggtga tgcagatgct gcttaacggc
     1381 ctctactaca tccacagaaa caagatcctg catagggaca tgaaggctgc taatgtgctt
     1441 atcactcgtg atggggtcct gaagctggca gactttgggc tggcccgggc cttcagcctg
     1501 gccaagaaca gccagcccaa ccgctacgcc aaccgtgtgg tgacactctg gtaccggccc
     1561 ccggagctgt tgctcgggga gcgggactac ggccccccca ttgacctgtg gggtgctggg
     1621 tgcatcatgg cagagatgtg gacccgcagc cccatcatgc agggcaacac ggagcagcac
     1681 caactcgccc tcatcagtca gctctgcggc tccatcaccc ctgaggtgtg gccaaacgtg
     1741 gacaactatg agctgtacga aaagctggag ctggtcaagg gccagaagcg gaaggtgaag
     1801 gacaggctga aggcctatgt gcgtgaccca tacgcactgg acctcatcga caagctgctg
     1861 gtgctggacc ctgcccagcg catcgacagc gatgacgccc tcaaccacga cttcttctgg
     1921 tccgacccca tgccctccga cctcaagggc atgctctcca cccacctgac gtccatgttc
     1981 gagtacttgg caccaccgcg ccggaagggc agccagatca cccagcagtc caccaaccag
     2041 agtcgcaatc ccgccaccac caaccagacg gagtttgagc gcgtcttctg aggatcccgg
     2101 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     2161 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     2221 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     2281 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     2341 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     2401 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     2461 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     2521 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     2581 tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc
     2641 ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac
     2701 tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt
     2761 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2821 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2881 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2941 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3001 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3061 aggcctaggc ttttgcaaaa agctcctcga ggaactgaaa aaccagaaag ttaattccct
     3121 atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     3181 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3241 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3301 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3361 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3421 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3481 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3541 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3601 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3661 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3721 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3781 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3841 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3901 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3961 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4021 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4081 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4141 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4201 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4261 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4321 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4381 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4441 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4501 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4561 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4621 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4681 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4741 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4801 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4861 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4921 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4981 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5041 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5101 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5161 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5221 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5281 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5341 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5401 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5461 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5521 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5581 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5641 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5701 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaattt
//
