LOCUS       sequence-324841-        6401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of CCNE1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6401)
  AUTHORS   Molina-Sanchez P, Ruiz de Galarreta M, Yao MA, Lindblad KE, 
            Bresnahan E, Bitterman E, Martin TC, Rubenstein T, Nie K, Golas J, 
            Choudhary S, Barcena-Varela M, Elmas A, Miguela V, Ding Y, Kan Z, 
            Grinspan LT, Huang KL, Parsons RE, Shields DJ, Rollins RA, Lujambio 
            A
  TITLE     Cooperation between distinct cancer driver genes underlies 
            inter-tumor heterogeneity in hepatocellular carcinoma.
  JOURNAL   Gastroenterology. 2020 Aug 16. pii: S0016-5085(20)35054-X. doi: 
            10.1053/j.gastro.2020.08.015.
  PUBMED    32814112
REFERENCE   2  (bases 1 to 6401)
  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: 
            "Gastroenterology. 2020 Aug 16. pii: S0016-5085(20)35054-X. doi: 
            10.1053/j.gastro.2020.08.015."
COMMENT     Sequence Label: pT3-EF1A-CCNE1
FEATURES             Location/Qualifiers
     source          1..6401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(385..405)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    3028..3149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3093..3112
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3780..3796)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3780..3796)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3793..3815)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3804..3820
                     /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(3828..3834,3835..3852,3853..3858))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3873..3894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4011..4028)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4182..4770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4262..4281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4941..5732,5733..5801))
                     /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"
     promoter        complement(5802..5906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5974..5992
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6030..6052)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6152..6171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6365..6387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6379..6396
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6380..6396
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aattcgagct cggtacccta cagttgaagt cggaagttta catacactta agttggagtc
       61 attaaaactc gtttttcaac tactccacaa atttcttgtt aacaaacaat agttttggca
      121 agtcagttag gacatctact ttgtgcatga cacaagtcat ttttccaaca attgtttaca
      181 gacagattat ttcacttata attcactgta tcacaattcc agtgggtcag aagtttacat
      241 acactaagtt gactgtgcct ttaaacagct tggaaaattc cagaaaatga tgtcatggct
      301 ttagaagctt ctgatagact aattgacatc atttgagtca attggaggtg tacctgtgga
      361 tgtatttcaa ggaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      421 gctccccagc aggcagaagt atgcaaagca tgcatttaat taaaattatc tctaaggcat
      481 gtgaactggc tgtcttggtt ttcatctgta cttcatctgc tacctctgtg acctgaaaca
      541 tatttataat tccattaagc tgtgcatatg atagatttat catatgtatt ttccttaaag
      601 gatttttgta agaactaatt gaattgatac ctgtaaagtc tttatcacac tacccaataa
      661 ataataaatc tctttgttca gctctctgtt tctataaata tgtaccagtt ttattgtttt
      721 tagtggtagt gattttattc tctttctata tatatacaca cacatgtgtg cattcataaa
      781 tatatacaat ttttatgaat aaaaaattat tagcaatcaa tattgaaaac cactgatttt
      841 tgtttatgtg agcaaacagc agattaaaag gaatttcaat tgcctgcagg agtcaatggg
      901 aaaaacccat tggagccaag tacactgact caatagggac tttccattgg gttttgccca
      961 gtacataagg tcaatagggg gtgagtcaac aggaaagtcc cattggagcc aagtacattg
     1021 agtcaatagg gactttccaa tgggttttgc ccagtacata aggtcaatgg gaggtaagcc
     1081 aatgggtttt tcccattact gacatgtata ctgagtcatt agggactttc caatgggttt
     1141 tgcccagtac ataaggtcaa taggggtgaa tcaacaggaa agtcccattg gagccaagta
     1201 cactgagtca atagggactt tccattgggt tttgcccagt acaaaaggtc aatagggggt
     1261 gagtcaatgg gtttttccca ttattggcac atacataagg tcaatagggg tgactagtgg
     1321 agaagagcat gcttgagggc tgagtgcccc tcagtgggca gagagcacat ggcccacagt
     1381 ccctgagaag ttggggggag gggtgggcaa ttgaactggt gcctagagaa ggtggggctt
     1441 gggtaaactg ggaaagtgat gtggtgtact ggctccacct ttttccccag ggtgggggag
     1501 aaccatatat aagtgcagta gtctctgtga acattcaagc ttctgccttc tccctcctgt
     1561 gagtttggta agtcactgac tgtctatgcc tgggaaaggg tgggcaggag atggggcagt
     1621 gcaggaaaag tggcactatg aaccctgcag ccctagacaa ttgtactaac cttcttctct
     1681 ttcctctcct gacaggttgg tgtacagtag cttccaagta ctaagatcta gtgcacaggg
     1741 cccaccatgg agctagctgg ggggatctca tgccgaggga gcgcagggag cgggatgcga
     1801 aggagcggga caccatgaag gaggacggcg gcgcggagtt ctcggctcgc tccaggaaga
     1861 ggaaggcaaa cgtgaccgtt tttttgcagg atccagatga agaaatggcc aaaatcgaca
     1921 ggacggcgag ggaccagtgt gggagccagc cttgggacaa taatgcagtc tgtgcagacc
     1981 cctgctccct gatccccaca cctgacaaag aagatgatga ccgggtttac ccaaactcaa
     2041 cgtgcaagcc tcggattatt gcaccatcca gaggctcccc gctgcctgta ctgagctggg
     2101 caaatagaga ggaagtctgg aaaatcatgt taaacaagga aaagacatac ttaagggatc
     2161 agcactttct tgagcaacac cctcttctgc agccaaaaat gcgagcaatt cttctggatt
     2221 ggttaatgga ggtgtgtgaa gtctataaac ttcacaggga gaccttttac ttggcacaag
     2281 atttctttga ccggtatatg gcgacacaag aaaatgttgt aaaaactctt ttacagctta
     2341 ttgggatttc atctttattt attgcagcca aacttgagga aatctatcct ccaaagttgc
     2401 accagtttgc gtatgtgaca gatggagctt gttcaggaga tgaaattctc accatggaat
     2461 taatgattat gaaactgttg gatctctgtg tcctggatgt tgactgcctt gaatttcctt
     2521 atggtatact tgctgcttcg gccttgtatc atttctcgtc atctgaattg atgcaaaagg
     2581 tttcagggta tcagtggtgc gacatagaga actgtgtcaa gtggatggtt ccatttgcca
     2641 tggttataag ggagacgggg agctcaaaac tgaagcactt caagggcgtc gctgatgaag
     2701 atgcacacaa catacagacc cacagagaca gcttggattt gctggacaaa gcccgagcaa
     2761 agaaagccat gttgtctgaa caaaataggg cttctcctct ccccagtggg ctcctcaccc
     2821 cgccacagag cggtaagaag cagagcagcg ggccggaaat ggcgtgaatc agtggtccag
     2881 gctctagttt tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata
     2941 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     3001 tttggcaagc tagctggcca gacatgataa gatacattga tgagtttgga caaaccacaa
     3061 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3121 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3181 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3241 tggaattgga gccccactgt gttcatctta cagatggaaa tactgacatt cagaggagtt
     3301 agttaacttg cctagcggcc gctgcattct agttgtggtt tgtccaaact catcaatgta
     3361 tcttatcatg tctggatccc cttgaaatac atccacaggt acacctccaa ttgactcaaa
     3421 tgatgtcaat tagtctatca gaagcttcta aagccatgac atcattttct ggaattttcc
     3481 aagctgttta aaggcacagt caacttagtg tatgtaaact tctgacccac tggaattgtg
     3541 atacagtgaa ttataagtga aataatctgt ctgtaaacaa ttgttggaaa aatgacttgt
     3601 gtcatgcaca aagtagatgt cctaactgac ttgccaaaac tattgtttgt taacaagaaa
     3661 tttgtggagt agttgaaaaa cgagttttaa tgactccaac ttaagtgtat gtaaacttcc
     3721 gacttcaact gtataggtct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg
     3781 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3841 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3901 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3961 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4021 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4081 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4141 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4201 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4261 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4321 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4381 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4441 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4501 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4561 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4621 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4681 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4741 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4801 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4861 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4921 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4981 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5041 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5101 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5161 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5221 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5281 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5341 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5401 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5461 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5521 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5581 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5641 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5701 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5761 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5821 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5881 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5941 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     6001 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     6061 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     6181 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     6241 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6301 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     6361 ttttcccagt cacgacgttg taaaacgacg gccagtgaat t
//
