LOCUS       sequence-194987-        7372 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 7372)
  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 7372)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..7372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    510..528
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    531..549
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    590..614
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      636..645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(642..644,645..647,648..1358)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(642..644,645..647,648..1358)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(687..708)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(948..967)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1295..1316
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1390..1414)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1467..1508
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(1546..1579)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1635..2223
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1688..1708)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2106..2117)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2226..2242)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2227..2243)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2752..2932
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2994..3582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3074..3093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3753..4544,4545..4613))
                     /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     4376..4395
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4614..4718)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4793..4812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4984..5363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5365..5563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5581..5761
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5808..5933
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6430..6663
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6848..6892
                     /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    7190..7307
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc tccctatcag
      541 tgatagagat cgtcgactag tccagtgtgg tggaattctg cagatatcaa caagtttgta
      601 caaaaaagca ggctttaaag gaaccgctac cggtcgccac catggtgagc aagggcgagg
      661 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      721 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      781 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
      841 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      901 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      961 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1021 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1081 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1141 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1201 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1261 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1321 ccgccgggat cactctcggc atggacgagc tgtacaagtc cggccggact cagatctcga
     1381 gatatctaga cccagctttc ttgtacaaag tggttgatat ccagcacagt ggcggccgct
     1441 cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg
     1501 attctacgcg taccggttag taatgacaat tcgtcgaggg acctaataac ttcgtatagc
     1561 atacattata cgaagttata catgtttaag ggttccggtt ccactaggta caattcgata
     1621 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1681 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1741 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     1801 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     1861 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     1921 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     1981 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     2041 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2101 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     2161 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     2221 cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat cacaagtagc
     2281 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
     2341 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     2401 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     2461 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac
     2521 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta
     2581 ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac
     2641 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac
     2701 agccgcctag catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc
     2761 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     2821 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     2881 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagcatgtga
     2941 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3001 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3061 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3121 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3181 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3241 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3301 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3361 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3421 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3481 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3541 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3601 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3661 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3721 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3781 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3841 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3901 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3961 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4021 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4081 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4141 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4201 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4261 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4321 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4381 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4441 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4501 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4561 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4621 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4681 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4741 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     4801 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     4861 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     4921 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     4981 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     5041 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     5101 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     5161 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     5221 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     5281 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     5341 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     5401 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     5461 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     5521 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact
     5581 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5641 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5701 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5761 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     5821 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     5881 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     5941 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     6001 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     6061 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     6121 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     6181 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     6241 aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg
     6301 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     6361 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     6421 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     6481 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     6541 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     6601 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     6661 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     6721 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     6781 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     6841 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     6901 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     6961 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     7021 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     7081 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     7141 ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa
     7201 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     7261 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     7321 gggacagcag agatccagtt tggttagtac cgggcccgct ctagcagttc cg
//
