LOCUS       sequence-324832-        6518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of CCNE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6518)
  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 6518)
  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-CCNE2
FEATURES             Location/Qualifiers
     source          1..6518
                     /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    3145..3266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3210..3229
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3897..3913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3897..3913)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3910..3932)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3921..3937
                     /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(3945..3951,3952..3969,3970..3975))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3990..4011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4128..4145)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4299..4887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4379..4398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5058..5849,5850..5918))
                     /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     5681..5700
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5919..6023)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6091..6109
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6147..6169)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6269..6288
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6482..6504
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6496..6513
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6497..6513
                     /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 tgtcaagacg aagtagccgt ttacaagcta
     1801 agcagcagcc ccagcccagc cagacggaat ccccccaaga agcccagata atccaggcca
     1861 agaagaggaa aactacccag gatgtcaaaa aaagaagaga ggaggtcacc aagaaacatc
     1921 agtatgaaat taggaattgt tggccacctg tattatctgg ggggatcagt ccttgcatta
     1981 tcattgaaac acctcacaaa gaaataggaa caagtgattt ctccagattt acaaattaca
     2041 gatttaaaaa tctttttatt aatccttcac ctttgcctga tttaagctgg ggatgttcaa
     2101 aagaagtctg gctaaacatg ttaaaaaagg agagcagata tgttcatgac aaacattttg
     2161 aagttctgca ttctgacttg gaaccacaga tgaggtccat acttctagac tggcttttag
     2221 aggtatgtga agtatacaca cttcataggg aaacatttta tcttgcacaa gacttttttg
     2281 atagatttat gttgacacaa aaggatataa ataaaaatat gcttcaactc attggaatta
     2341 cctcattatt cattgcttcc aaacttgagg aaatctatgc tcctaaactc caagagtttg
     2401 cttacgtcac tgatggtgct tgcagtgaag aggatatctt aaggatggaa ctcattatat
     2461 taaaggcttt aaaatgggaa ctttgtcctg taacaatcat ctcctggcta aatctctttc
     2521 tccaagttga tgctcttaaa gatgctccta aagttcttct acctcagtat tctcaggaaa
     2581 cattcattca aatagctcag cttttagatc tgtgtattct agccattgat tcattagagt
     2641 tccagtacag aatactgact gctgctgcct tgtgccattt tacctccatt gaagtggtta
     2701 agaaagcctc aggtttggag tgggacagta tttcagaatg tgtagattgg atggtacctt
     2761 ttgtcaatgt agtaaaaagt actagtccag tgaagctgaa gacttttaag aagattccta
     2821 tggaagacag acataatatc cagacacata caaactattt ggctatgctg gaggaagtaa
     2881 attacataaa caccttcaga aaagggggac agttgtcacc agtgtgcaat ggaggcatta
     2941 tgacaccacc gaagagcact gaaaaaccac caggaaaaca ctaaatcagt ggtccaggct
     3001 ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa
     3061 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     3121 ggcaagctag ctggccagac atgataagat acattgatga gtttggacaa accacaacta
     3181 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3241 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3301 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
     3361 aattggagcc ccactgtgtt catcttacag atggaaatac tgacattcag aggagttagt
     3421 taacttgcct agcggccgct gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3481 tatcatgtct ggatcccctt gaaatacatc cacaggtaca cctccaattg actcaaatga
     3541 tgtcaattag tctatcagaa gcttctaaag ccatgacatc attttctgga attttccaag
     3601 ctgtttaaag gcacagtcaa cttagtgtat gtaaacttct gacccactgg aattgtgata
     3661 cagtgaatta taagtgaaat aatctgtctg taaacaattg ttggaaaaat gacttgtgtc
     3721 atgcacaaag tagatgtcct aactgacttg ccaaaactat tgtttgttaa caagaaattt
     3781 gtggagtagt tgaaaaacga gttttaatga ctccaactta agtgtatgta aacttccgac
     3841 ttcaactgta taggtctaga gtcgacctgc aggcatgcaa gcttggcgta atcatggtca
     3901 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3961 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4021 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4081 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4141 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4201 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4261 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4321 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4381 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4441 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4501 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4561 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4621 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4681 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4741 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4801 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4861 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4921 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4981 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5041 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5101 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5161 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5221 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5281 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5341 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5401 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5461 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5521 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5581 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5641 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5701 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5761 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5821 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5881 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5941 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6001 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6061 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     6121 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     6181 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6241 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     6301 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     6361 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     6421 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     6481 tcccagtcac gacgttgtaa aacgacggcc agtgaatt
//
