LOCUS       sequence-110663-        8263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8263)
  AUTHORS   Blelloch R, Venere M, Yen J, Ramalho-Santos M
  TITLE     Generation of induced pluripotent stem cells in the absence of drug 
            selection.
  JOURNAL   Cell Stem Cell. 2007 Sep 13;1(3):245-7. doi: 
            10.1016/j.stem.2007.08.008. Epub 2007 Sep 6.
  PUBMED    18371358
REFERENCE   2  (bases 1 to 8263)
  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: 
            "10.1016/j.stem.2007.08.008"; journalName: "Cell Stem Cell"; date: 
            "2007-09-13- 13"; volume: "1"; issue: "3"; pages: "245-7"
COMMENT     Sequence Label: pLOVE empty vector
FEATURES             Location/Qualifiers
     source          1..8263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        247..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        628..826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             844..1024
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1071..1196
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1695..1928
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2113..2157
                     /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    2455..2572
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2638..2941
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2942..3145
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    3147..3165
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3168..3186
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3227..3351
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3376..3381,3382..3399,3400..3406)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3460..4119
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3526..3545)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4461..4766
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4645..4664
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4807..4931)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4984..5025
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(5063..5096)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5152..5740
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5205..5225)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5623..5634)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5743..5759)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5744..5760)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6269..6449
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6511..7099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6591..6610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7270..8061,8062..8130))
                     /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     7893..7912
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8131..8235)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcttaagcgg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct
     1561 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     1621 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     1681 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     1741 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     1801 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     1861 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     1921 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     1981 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     2041 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     2101 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     2161 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     2221 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     2281 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     2341 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     2401 ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa
     2461 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     2521 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     2581 ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctagc agttccgcgt
     2641 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     2701 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     2761 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     2821 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     2881 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     2941 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     3001 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     3061 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     3121 gtgggaggtc tatataagca gagctctccc tatcagtgat agagatctcc ctatcagtga
     3181 tagagatcgt cgactagtcc agtgtggtgg aattctgcag atatcaacaa gtttgtacaa
     3241 aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca
     3301 taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat ggcggccgca
     3361 ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg gattttgagt
     3421 taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     3481 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     3541 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     3601 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     3661 aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt
     3721 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     3781 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     3841 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     3901 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     3961 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     4021 gcgattcagg ttcatcatgc cgtctgtgat ggcttccatg tcggcagaat gcttaatgaa
     4081 ttacaacagt actgcgatga gtggcagggc ggggcgtaaa gatctggatc cggcttacta
     4141 aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact
     4201 gatatgtata cccgaagtat gtcaaaaaga ggtgtgctat gaagcagcgt attacagtga
     4261 cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct
     4321 ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga
     4381 aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga
     4441 cgagaacagg gactggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt
     4501 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga
     4561 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     4621 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     4681 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     4741 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag
     4801 gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc
     4861 aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg
     4921 tacaaagtgg ttgatatcca gcacagtggc ggccgctcga gtctagaggg cccgcggttc
     4981 gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa
     5041 tgacaattcg tcgagggacc taataacttc gtatagcata cattatacga agttatacat
     5101 gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga taatcaacct
     5161 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     5221 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     5281 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     5341 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     5401 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     5461 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     5521 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     5581 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     5641 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     5701 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
     5761 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     5821 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     5881 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     5941 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     6001 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     6061 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     6121 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     6181 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     6241 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     6301 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     6361 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     6421 cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag
     6481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7021 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7081 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7141 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7201 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7261 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7321 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7381 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7441 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7501 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7561 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7621 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7681 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7741 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7801 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7861 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7921 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7981 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8041 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8101 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8161 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8221 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gac
//
