LOCUS       Exported                7967 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Overexpression of Tert.
ACCESSION   .
VERSION     .
KEYWORDS    pT3-EF1A-mTert
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7967)
  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 7967)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7967
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(385..405)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    4594..4715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4659..4678
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5346..5362)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5346..5362)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5359..5381)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5370..5386
                     /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(5394..5424)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5439..5460
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5577..5594)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5748..6336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5828..5847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6507..7367)
                     /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     7130..7149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7368..7472)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7540..7558
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7596..7618)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7718..7737
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7931..7953
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7945..7962
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7946..7962
                     /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 tgacccgcgc tcctcgttgc cccgcggtgc
     1801 gctctctgct gcgcagccga taccgggagg tgtggccgct ggcaaccttt gtgcggcgcc
     1861 tggggcccga gggcaggcgg cttgtgcaac ccggggaccc gaagatctac cgcactttgg
     1921 ttgcccaatg cctagtgtgc atgcactggg gctcacagcc tccacctgcc gacctttcct
     1981 tccaccaggt gtcatccctg aaagagctgg tggccagggt tgtgcagaga ctctgcgagc
     2041 gcaacgagag aaacgtgctg gcttttggct ttgagctgct taacgaggcc agaggcgggc
     2101 ctcccatggc cttcactagt agcgtgcgta gctacttgcc caacactgtt attgagaccc
     2161 tgcgtgtcag tggtgcatgg atgctactgt tgagccgagt gggcgacgac ctgctggtct
     2221 acctgctggc acactgtgct ctttatcttc tggtgccccc cagctgtgcc taccagggga
     2281 gatggccaag agcgtctaaa cccctcattc ctactcagca acctccagcc taacttgact
     2341 ggggccagga gactggtgga gatcatcttt ctgggctcaa ggcctaggac atcaggacca
     2401 ctctgcagga cacaccgtct atcgcgtcga tactggcaga tgcggcccct gttccaacag
     2461 ctgctggtga accatgcaga gtgccaatat gtcagactcc tcaggtcaca ttgcaggttt
     2521 cgaacagcaa accaacaggt gacagatgcc ttgaacacca gcccaccgca cctcatggat
     2581 ttgctccgcc tgcacagcag tccctggcag ggaaggaccg tgtccccgct gcagagcacc
     2641 gtctgaggga gaggatcctg gctacgttcc tgttctggct gatggacaca tacgtggtac
     2701 agctgcttag gtcattcttt tacatcacag agagcacatt ccagaagaac aggctcttct
     2761 tctaccgtaa gagtgtgtgg agcaagctgc agagcattgg agtcaggcaa caccttgaga
     2821 gagtgcggct acgggagctg tcacaagagg aggtcaggca tcaccaggac acctggctag
     2881 ccatgcccat ctgcagactg cgcttcatcc ccaagcccaa cggcctgcgg cccattgtga
     2941 acatgagtta tagcatgggt accagagctt tgggcagaag gaagcaggcc cagcatttca
     3001 cccagcgtct caagactctc ttcagcatgc tcaactatga gcggacaaaa catcctcacc
     3061 ttatggggtc ttctgtactg ggtatgaatg acatctacag gacctggcgg gcctttgtgc
     3121 tgcgtgtgcg tgctctggac cagacaccca ggatgtactt tgttaaggca gatgtgaccg
     3181 gggcctatga tgccatcccc cagggtaagc tggtggaggt tgttgccaat atgatcaggc
     3241 actcggagag cacgtactgt atccgccagt atgcagtggt ccggagagat agccaaggcc
     3301 aagtccacaa gtcctttagg agacaggtca ccaccctctc tgacctccag ccatacatgg
     3361 gccagttcct taagcatctg caggattcag atgccagtgc actgaggaac tccgttgtca
     3421 tcgagcagag catctctatg aatgagagca gcagcagcct gtttgacttc ttcctgcact
     3481 tcctgcgtca cagtgtcgta aagattggtg acaggtgcta tacgcagtgc cagggcatcc
     3541 cccagggctc cagcctatcc accctgctct gcagtctgtg tttcggagac atggagaaca
     3601 agctgtttgc tgaggtgcag cgggatgggt tgcttttacg ttttgttgat gactttctgt
     3661 tggtgacgcc tcacttggac caagcaaaaa ccttcctcag caccctggtc catggcgttc
     3721 ctgagtatgg gtgcatgata aacttgcaga agacagtggt gaacttccct gtggagcctg
     3781 gtaccctggg tggtgcagct ccataccagc tgcctgctca ctgcctgttt ccctggtgtg
     3841 gcttgctgct ggacactcag actttggagg tgttctgtga ctactcaggt tatgcccaga
     3901 cctcaattaa gacgagcctc accttccaga gtgtcttcaa agctgggaag accatgcgga
     3961 acaagctcct gtcggtcttg cggttgaagt gtcacggtct atttctagac ttgcaggtga
     4021 acagcctcca gacagtctgc atcaatatat acaagatctt cctgcttcag gcctacaggt
     4081 tccatgcatg tgtgattcag cttccctttg accagcgtgt taggaagaac ctcacattct
     4141 ttctgggcat catctccagc caagcatcct gctgctatgc tatcctgaag gtcaagaatc
     4201 caggaatgac actaaaggcc tctggctcct ttcctcctga agccgcacat tggctctgct
     4261 accaggcctt cctgctcaag ctggctgctc attctgtcat ctacaaatgt ctcctgggac
     4321 ctctgaggac agcccaaaaa ctgctgtgcc ggaagctccc agaggcgaca atgaccatcc
     4381 ttaaagctgc agctgaccca gccctaagca cagactttca gaccattttg gacatcagtg
     4441 gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca
     4501 tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     4561 tgtaggtttg gcaagctagc tggccagaca tgataagata cattgatgag tttggacaaa
     4621 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     4681 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     4741 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     4801 gtggtatgga attggagccc cactgtgttc atcttacaga tggaaatact gacattcaga
     4861 ggagttagtt aacttgccta gcggccgctg cattctagtt gtggtttgtc caaactcatc
     4921 aatgtatctt atcatgtctg gatccccttg aaatacatcc acaggtacac ctccaattga
     4981 ctcaaatgat gtcaattagt ctatcagaag cttctaaagc catgacatca ttttctggaa
     5041 ttttccaagc tgtttaaagg cacagtcaac ttagtgtatg taaacttctg acccactgga
     5101 attgtgatac agtgaattat aagtgaaata atctgtctgt aaacaattgt tggaaaaatg
     5161 acttgtgtca tgcacaaagt agatgtccta actgacttgc caaaactatt gtttgttaac
     5221 aagaaatttg tggagtagtt gaaaaacgag ttttaatgac tccaacttaa gtgtatgtaa
     5281 acttccgact tcaactgtat aggtctagag tcgacctgca ggcatgcaag cttggcgtaa
     5341 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5401 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5461 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5521 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5581 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5641 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5701 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5761 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5821 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5881 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5941 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6001 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6061 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6181 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6421 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6481 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6541 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6601 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6661 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6721 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6781 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6841 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6901 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6961 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7021 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7081 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7141 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7201 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7261 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7321 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7381 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7441 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7501 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7561 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     7621 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     7681 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     7741 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     7801 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     7861 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     7921 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaatt
//
