LOCUS       sequence-193283-        6356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of thymidylate synthetase .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6356)
  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 6356)
  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-TYMS
FEATURES             Location/Qualifiers
     source          1..6356
                     /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(1713..1730)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1719..1943
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1989..2417
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2076..2095)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2286..2307
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2426..2446)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2431..2760
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2611..2746
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2673..2692
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2827..3621
                     /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(2881..2900)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3491..3510
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3795..3916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3832..3851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3886..3905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3965..3981)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3965..3981)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3978..4000)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3989..4005
                     /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(4013..4019,4020..4037,4038..4043))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4058..4079
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4196..4213)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4367..4955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4447..4466)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5126..5917,5918..5986))
                     /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     5749..5768
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5987..6091)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6166..6185)
                     /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 atcatgcctg tggccggctc ggagctgccg cgccggccct tgccccccgc
      781 cgcacaggag cgggacgccg agccgcgtcc gccgcacggg gagctgcagt acctggggca
      841 gatccaacac atcctccgct gcggcgtcag gaaggacgac cgcacgggca ccggcaccct
      901 gtcggtattc ggcatgcagg cgcgctacag cctgagagat gaattccctc tgctgacaac
      961 caaacgtgtg ttctggaagg gtgttttgga ggagttgctg tggtttatca agggatccac
     1021 aaatgctaaa gagctgtctt ccaagggagt gaaaatctgg gatgccaatg gatcccgaga
     1081 ctttttggac agcctgggat tctccaccag agaagaaggg gacttgggcc cagtttatgg
     1141 cttccagtgg aggcattttg gggcagaata cagagatatg gaatcagatt attcaggaca
     1201 gggagttgac caactgcaaa gagtgattga caccatcaaa accaaccctg acgacagaag
     1261 aatcatcatg tgcgcttgga atccaagaga tcttcctctg atggcgctgc ctccatgcca
     1321 tgccctctgc cagttctatg tggtgaacag tgagctgtcc tgccagctgt accagagatc
     1381 gggagacatg ggcctcggtg tgcctttcaa catcgccagc tacgccctgc tcacgtacat
     1441 gattgcgcac atcacgggcc tgaagccagg tgactttata cacactttgg gagatgcaca
     1501 tatttacctg aatcacatcg agccactgaa aattcagctt cagcgagaac ccagaccttt
     1561 cccaaagctc aggattcttc gaaaagttga gaaaattgat gacttcaaag ctgaagactt
     1621 tcagattgaa gggtacaatc cgcatccaac tattaaaatg gaaatggctg tttagctcga
     1681 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     1741 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1801 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1861 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     1921 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     1981 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2041 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2101 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2161 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2221 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2281 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2341 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2401 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2461 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     2521 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2581 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     2641 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     2701 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     2761 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     2821 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2881 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2941 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3001 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3061 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3121 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3181 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3241 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3301 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     3361 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3421 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3481 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3541 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3601 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3661 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3721 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3781 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3841 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3901 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     3961 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4021 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4081 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4141 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4201 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4261 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4321 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4381 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4441 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4501 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4561 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4621 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4681 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4741 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4801 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4861 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4921 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4981 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5041 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5101 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5161 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5221 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5281 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5341 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5401 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5461 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5521 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5581 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5641 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5701 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5761 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5821 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5881 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5941 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6001 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6061 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     6181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     6241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     6301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
