LOCUS       Exported                8857 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Dox-inducible lentiviral vector expressing SV40 large T antigen for 
            immortalizing cells.
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE2-TetOminiCMV-SV40LT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8857)
  AUTHORS   Velychko S, Adachi K, Kim KP, Hou Y, MacCarthy CM, Wu G, Scholer HR
  TITLE     Excluding Oct4 from Yamanaka Cocktail Unleashes the Developmental 
            Potential of iPSCs.
  JOURNAL   Cell Stem Cell. 2019 Oct 30. pii: S1934-5909(19)30423-0. doi: 
            10.1016/j.stem.2019.10.002.
  PUBMED    31708402
REFERENCE   2  (bases 1 to 8857)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8857
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2251..2521
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2554..2592
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2589..2613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     2684..2700
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2684..2700
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             join(2778..3023,3370..5250)
                     /codon_start=1
                     /product="SV40 (simian virus 40) large T antigen"
                     /label=large T antigen
                     /translation="MDKVLNREESLQLMDLLGLERSAWGNIPLMRKAYLKKCKEFHPDK
                     GGDEEKMKKMNTLYKKMEDGVKYAHQPDFGGFWDATEIPTYGTDEWEQWWNAFNEENLF
                     CSEEMPSSDDEATADSQHSTPPKKKRKVEDPKDFPSELLSFLSHAVFSNRTLACFAIYT
                     TKEKAALLYKKIMEKYSVTFISRHNSYNHNILFFLTPHRHRVSAINNYAQKLCTFSFLI
                     CKGVNKEYLMYSALTRDPFSVIEESLPGGLKEHDFNPEEAEETKQVSWKLVTEYAMETK
                     CDDVLLLLGMYLEFQYSFEMCLKCIKKEQPSHYKYHEKHYANAAIFADSKNQKTICQQA
                     VDTVLAKKRVDSLQLTREQMLTNRFNDLLDRMDIMFGSTGSADIEEWMAGVAWLHCLLP
                     KMDSVVYDFLKCMVYNIPKKRYWLFKGPIDSGKTTLAAALLELCGGKALNVNLPLDRLN
                     FELGVAIDQFLVVFEDVKGTGGESRDLPSGQGINNLDNLRDYLDGSVKVNLEKKHLNKR
                     TQIFPPGIVTMNEYSVPKTLQARFVKQIDFRPKDYLKHCLERSEFLLEKRIIQSGIALL
                     LMLIWYRPVAEFAQSIQSRIVEWKERLDKEFSLSVYQKMKFNVAMGIGVLDWLRNSDDD
                     DEDSQENADKNEDGGEKNMEDSGHETGIDSQSQGSFQAPQSSQSVHDHNQPYHICRGFT
                     CFKKPPTPPPEPET"
     intron          3024..3369
                     /label=large T intron
                     /note="SV40 (simian virus 40) large T antigen intron"
     misc_feature    5299..5887
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5352..5372)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5770..5781)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6035..6215
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6262..6280)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6348..6452
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6453..7313
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6671..6690)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7484..8072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7973..7992
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8226..8243
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8335..8354
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    8483..8504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8519..8549
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8557..8573
                     /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)."
     primer_bind     8562..8584
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8581..8597
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8581..8597
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8647..8666
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcca
     2221 caccctaact gacacactcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
     2281 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
     2341 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     2401 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     2461 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     2521 aaaagtgaaa gtcgagctcg gtacccgggt cgaggtaggc gtgtacggtg ggaggcctat
     2581 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     2641 gacctccata gaagacaccg ggaccgatcc agcctgcggc cgctctagaa ctagtggatc
     2701 cgcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2761 caaaaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg cagctaatgg
     2821 accttctagg tcttgaaagg agtgcctggg ggaatattcc tctgatgaga aaggcatatt
     2881 taaaaaaatg caaggagttt catcctgata aaggaggaga tgaagaaaaa atgaagaaaa
     2941 tgaatactct gtacaagaaa atggaagatg gagtaaaata tgctcatcaa cctgactttg
     3001 gaggcttctg ggatgcaact gaggtatttg cttcttcctt aaatcctggt gttgatgcaa
     3061 tatactgcaa acaatggcct gagtgtgtaa agaaaatgtc tactaactgc atatgcttgc
     3121 tgtgcttact gaggatgaag catgaaaata gaaaattata caggaaagat ccacttgtgt
     3181 gggttgattg ttactgcttc gattgcttta gaatgtggtt tggacttgat ctttgtgaag
     3241 gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct
     3301 aaggtaaata taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt
     3361 gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat
     3421 gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac
     3481 tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct
     3541 tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt
     3601 gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat
     3661 tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt ttttcttact
     3721 ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg tacctttagc
     3781 tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt gactagagat
     3841 ccattttctg ttattgagga aagtttgcca ggtgggttaa aggagcatga ttttaatcca
     3901 gaagaagcag aggaaactaa acaagtgtcc tggaagcttg taacagagta tgcaatggaa
     3961 acaaaatgtg atgatgtgtt gttattgctt gggatgtact tggaatttca gtacagtttt
     4021 gaaatgtgtt taaaatgtat taaaaaagaa cagcccagcc actataagta ccatgaaaag
     4081 cattatgcaa atgctgctat atttgctgac agcaaaaacc aaaaaaccat atgccaacag
     4141 gctgttgata ctgttttagc taaaaagcgg gttgatagcc tacaattaac tagagaacaa
     4201 atgttaacaa acagatttaa tgatcttttg gataggatgg atataatgtt tggttctaca
     4261 ggctctgctg acatagaaga atggatggct ggagttgctt ggctacactg tttgttgccc
     4321 aaaatggatt cagtggtgta tgacttttta aaatgcatgg tgtacaacat tcctaaaaaa
     4381 agatactggc tgtttaaagg accaattgat agtggtaaaa ctacattagc agctgctttg
     4441 cttgaattat gtggggggaa agctttaaat gttaatttgc ccttggacag gctgaacttt
     4501 gagctaggag tagctattga ccagttttta gtagtttttg aggatgtaaa gggcactgga
     4561 ggggagtcca gagatttgcc ttcaggtcag ggaattaata acctggacaa tttaagggat
     4621 tatttggatg gcagtgttaa ggtaaactta gaaaagaaac acctaaataa aagaactcaa
     4681 atatttcccc ctggaatagt caccatgaat gagtacagtg tgcctaaaac actgcaggcc
     4741 agatttgtaa aacaaataga ttttaggccc aaagattatt taaagcattg cctggaacgc
     4801 agtgagtttt tgttagaaaa gagaataatt caaagtggca ttgctttgct tcttatgtta
     4861 atttggtaca gacctgtggc tgagtttgct caaagtattc agagcagaat tgtggagtgg
     4921 aaagagagat tggacaaaga gtttagtttg tcagtgtatc aaaaaatgaa gtttaatgtg
     4981 gctatgggaa ttggagtttt agattggcta agaaacagtg atgatgatga tgaagacagc
     5041 caggaaaatg ctgataaaaa tgaagatggt ggggagaaga acatggaaga ctcagggcat
     5101 gaaacaggca ttgattcaca gtcccaaggc tcatttcagg cccctcagtc ctcacagtct
     5161 gttcatgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     5221 cccacacctc cccctgaacc tgaaacataa aggatcctta agctagcgaa ttcgatatca
     5281 agcttatcga tagatcctaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5341 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     5401 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5461 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     5521 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     5581 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     5641 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     5701 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5761 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5821 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5881 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     5941 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatcacct
     6001 gcaggacagg cgcgccctgc tttttgcttg tactgggtct ctctggttag accagatctg
     6061 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     6121 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     6181 cagacccttt tagtcagtgt ggaaaatctc tagcacccgg gcgattaagg aaagggctag
     6241 atcattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     6301 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     6361 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     6421 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6481 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6541 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6601 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6661 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     6721 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6781 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6841 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6901 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6961 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     7021 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     7081 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7141 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     7201 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     7261 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     7321 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     7381 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     7441 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7501 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7561 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7621 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7681 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7741 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7801 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7861 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7921 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7981 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     8041 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     8101 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     8161 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     8221 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     8281 ccgcgcgttg gccgattcat taatgcagca agctcatggc tgactaattt tttttattta
     8341 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     8401 tggaggccta ggcttttgca aaaagctccc cgtggcacga caggtttccc gactggaaag
     8461 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     8521 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     8581 caggaaacag ctatgacatg attacgaatt tcacaaataa agcatttttt tcactgcatt
     8641 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa
     8701 ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt
     8761 acctgtggtt tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa
     8821 attgtatttg ttaaatatgt actacaaact tagtagt
//
