LOCUS       sequence-193284-        6053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of deoxythymidylate kinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6053)
  AUTHORS   Yeh HW, Lee SS, Chang CY, Hu CM, Jou YS
  TITLE     Pyrimidine metabolic rate limiting enzymes in poorly-differentiated 
            hepatocellular carcinoma are signature genes of cancer stemness and 
            associated with poor prognosis.
  JOURNAL   Oncotarget. 2017 Sep 8;8(44):77734-77751. doi: 
            10.18632/oncotarget.20774. eCollection 2017 Sep 29.
  PUBMED    29100421
REFERENCE   2  (bases 1 to 6053)
  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"; doi: 
            "10.18632/oncotarget.20774"; journalName: "Oncotarget"; date: 
            "2017-09-8- 8"; volume: "8"; issue: "44"; pages: "77734-77751"
COMMENT     Sequence Label: pcDNA3.1-DTYMK
FEATURES             Location/Qualifiers
     source          1..6053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1410..1427)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1416..1640
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1686..2114
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1773..1792)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1983..2004
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2123..2143)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2128..2457
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2308..2443
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2370..2389
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2524..3318
                     /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(2578..2597)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3188..3207
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3492..3613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3529..3548)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3583..3602
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3662..3678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3662..3678)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3675..3697)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3686..3702
                     /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(3710..3716,3717..3734,3735..3740))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3755..3776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3893..3910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4064..4652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4144..4163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4823..5614,5615..5683))
                     /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     5446..5465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5684..5788)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5863..5882)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      721 ttctgcagat atcatggcgg cccggcgcgg ggctctcata gtgctggagg gcgtggaccg
      781 cgccgggaag agcacgcaga gccgcaagct ggtggaagcg ctgtgcgccg cgggccaccg
      841 cgccgaactg ctccggttcc cggaaagatc aactgaaatc ggcaaacttc tgagttccta
      901 cttgcaaaag aaaagtgacg tggaggatca ctcggtgcac ctgctttttt ctgcaaatcg
      961 ctgggaacaa gtgccgttaa ttaaggaaaa gttgagccag ggcgtgaccc tcgtcgtgga
     1021 cagatacgca ttttctggtg tggccttcac cggtgccaag gagaattttt ccctagattg
     1081 gtgtaaacag ccagacgtgg gccttcccaa acccgacctg gtcctgttcc tccagttaca
     1141 gctggcggat gctgccaagc ggggagcgtt tggccatgag cgctatgaga acggggcttt
     1201 ccaggagcgg gcgctccggt gtttccacca gctcatgaaa gacacgactt tgaactggaa
     1261 gatggtggat gcttccaaaa gcatcgaagc tgtccatgag gacatccgcg tgctctctga
     1321 ggacgccatc cgcactgcca cagagaagcc gctgggggag ctatggaagt gactcgagtc
     1381 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     1441 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1501 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1561 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     1621 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     1681 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1741 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1801 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1861 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1921 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1981 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2041 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2101 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2161 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     2221 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2281 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2341 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     2401 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     2461 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     2521 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2581 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2641 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2701 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2761 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2821 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     2881 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     2941 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3001 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3061 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3121 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3181 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3241 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3301 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     3361 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3421 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3481 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3541 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3601 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     3661 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3721 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3781 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3841 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3901 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3961 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4021 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4081 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4141 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4201 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4261 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4321 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4381 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4441 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4501 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4561 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4621 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4681 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4741 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4801 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4861 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4921 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4981 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5041 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5101 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5161 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5221 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5281 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5341 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5401 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5461 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5521 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5581 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5641 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5701 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5761 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     5821 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     5881 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     5941 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     6001 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gtt
//
