LOCUS       sequence-364657-        7765 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 7765)
  AUTHORS   Kita-Matsuo H, Barcova M, Prigozhina N, Salomonis N, Wei K, Jacot 
            JG, Nelson B, Spiering S, Haverslag R, Kim C, Talantova M, Bajpai R,
            Calzolari D, Terskikh A, McCulloch AD, Price JH, Conklin BR, Chen 
            HS, Mercola M
  TITLE     Lentiviral vectors and protocols for creation of stable hESC lines 
            for fluorescent tracking and drug resistance selection of 
            cardiomyocytes.
  JOURNAL   PLoS ONE. 2009  . 4(4):e5046.
  PUBMED    19352491
REFERENCE   2  (bases 1 to 7765)
  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: "PLoS ONE. 
            2009  . 4(4):e5046."
COMMENT     Sequence Label: PGK-H2BeGFP
FEATURES             Location/Qualifiers
     source          1..7765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3..22)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4..22)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(32..49)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(32..48)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(41..63)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      190..645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(277..296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     487..508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        671..775
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(776..844,845..1636)
                     /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     complement(994..1013)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1807..2395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2296..2315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2549..2566
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2683..2704
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2719..2724,2725..2742,2743..2749)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2757..2773
                     /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     2762..2784
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2781..2797
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2781..2797
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2816..2836
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2818..2836
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2864..3090
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3091..3271
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3318..3443
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3936..4169
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4354..4398
                     /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    4665..4782
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4828..5338
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     5257..5275
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             5347..5724
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     regulatory      5731..5740
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5737..5739,5740..5742,5743..6456)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5737..5739,5740..5742,5743..6453)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5782..5803)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6043..6062)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6390..6411
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6472..7060
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6525..6545)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6943..6954)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7147..7380
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7452..7573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7489..7508)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7543..7562
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7613..7748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
       61 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
      121 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
      181 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
      241 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
      301 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
      361 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
      421 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
      481 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
      541 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
      601 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
      661 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
      721 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      781 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
      841 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
      901 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
      961 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     1021 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1081 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1141 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1201 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1261 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1321 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1381 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1441 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1501 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1561 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1621 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1681 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1741 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1801 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1861 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1921 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     1981 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2041 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2101 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2161 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2221 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2281 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2341 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2401 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2461 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2521 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2581 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     2641 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     2701 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     2761 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     2821 taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata
     2881 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
     2941 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
     3001 caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt
     3061 gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc
     3121 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     3181 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     3241 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     3301 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     3361 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     3421 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     3481 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     3541 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     3601 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     3661 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     3721 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     3781 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     3841 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     3901 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     3961 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     4021 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     4081 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     4141 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     4201 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     4261 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     4321 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     4381 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     4441 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     4501 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     4561 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     4621 agatccattc gattagtgaa cggatctcga cggtatcggt taacttttaa aagaaaaggg
     4681 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     4741 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttatcgatca cgagactagc
     4801 ctcgagaagc ttgatatcga attccacggg gttggggttg cgccttttcc aaggcagccc
     4861 tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca gcggcgccga
     4921 ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct tcgccgctac
     4981 ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag gttccttgcg
     5041 gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta ccctcgcaga
     5101 cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg ccaatagcgg
     5161 ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg aggcggggtg
     5221 tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct gcaagcctcc
     5281 ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct ctccccaggg
     5341 ggatccatgc cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg
     5401 gtgactaagg cgcagaagaa agacggcaag aagcgcaagc gcagccgcaa ggagagctat
     5461 tccatctatg tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag
     5521 gccatgggca tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct
     5581 tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc
     5641 gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc
     5701 gtcaccaagt acaccagcgc taaaccggtc gccaccatgg tgagcaaggg cgaggagctg
     5761 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     5821 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     5881 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     5941 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     6001 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     6061 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     6121 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     6181 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     6241 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     6301 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     6361 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     6421 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgtcga caatcaacct
     6481 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     6541 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     6601 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     6661 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     6721 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     6781 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     6841 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     6901 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     6961 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     7021 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac
     7081 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     7141 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     7201 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     7261 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     7321 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     7381 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     7441 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     7501 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     7561 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     7621 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     7681 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     7741 tggaggccta gggacgtacc caatt
//
