LOCUS       sequence-447568-        7756 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 7756)
  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 7756)
  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-H2BmCherry
FEATURES             Location/Qualifiers
     source          1..7756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     296..315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     549..566
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    683..704
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(719..724,725..742,743..749)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    757..773
                     /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     762..784
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     781..797
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     781..797
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     816..836
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        818..836
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        864..1090
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1091..1271
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1318..1443
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1936..2169
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2354..2398
                     /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    2665..2782
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2828..3338
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3257..3275
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3353..3730
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     CDS             3737..4447
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3882..3900)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4150..4169
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4463..5051
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4516..4536)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4934..4945)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5138..5371
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5443..5564
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5480..5499)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5534..5553
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5604..5739
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5759..5778)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5760..5778)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5788..5805)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5788..5804)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5797..5819)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5946..6401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6033..6052)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6243..6264
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6427..6531
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6532..6600,6601..7392)
                     /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(6750..6769)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7563..7756,1..395)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
       61 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      121 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      181 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      241 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      301 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      361 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      421 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      481 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      541 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      601 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      661 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      721 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      781 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
      841 aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat
      901 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt
      961 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg
     1021 gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg
     1081 atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     1141 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     1201 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     1261 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
     1321 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     1381 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     1441 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     1501 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     1561 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     1621 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     1681 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     1741 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
     1801 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     1861 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     1921 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     1981 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     2041 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     2101 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     2161 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     2221 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     2281 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     2341 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     2401 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     2461 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     2521 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     2581 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     2641 cggatctcga cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc
     2701 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     2761 aattacaaaa attcaaaatt ttatcgatca cgagactagc ctcgagaagc ttgatatcga
     2821 attccacggg gttggggttg cgccttttcc aaggcagccc tgggtttgcg cagggacgcg
     2881 gctgctctgg gcgtggttcc gggaaacgca gcggcgccga ccctgggtct cgcacattct
     2941 tcacgtccgt tcgcagcgtc acccggatct tcgccgctac ccttgtgggc cccccggcga
     3001 cgcttcctgc tccgccccta agtcgggaag gttccttgcg gttcgcggcg tgccggacgt
     3061 gacaaacgga agccgcacgt ctcactagta ccctcgcaga cggacagcgc cagggagcaa
     3121 tggcagcgcg ccgaccgcga tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg
     3181 agagcagcgg ccgggaaggg gcggtgcggg aggcggggtg tggggcggta gtgtgggccc
     3241 tgttcctgcc cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg
     3301 gctccctcgt tgaccgaatc accgacctct ctccccaggg ggatccgcca ccatgccaga
     3361 gccagcgaag tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca
     3421 gaagaaagac ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta
     3481 caaggttctg aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat
     3541 gaattcgttt gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca
     3601 ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct
     3661 gcctggggag ttggccaagc acgccgtgtc cgagggtact aaggccgtca ccaagtacac
     3721 cagcgctaag gatccgatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     3781 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     3841 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     3901 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     3961 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     4021 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     4081 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     4141 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     4201 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     4261 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     4321 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     4381 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     4441 caagtaaagc ggccgcgtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     4501 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4561 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4621 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4681 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4741 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4801 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     4861 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4921 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4981 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5041 cgcctccccg cctggaattc gagctcggta cctttaagac caatgactta caaggcagct
     5101 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     5161 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     5221 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     5281 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     5341 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     5401 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     5461 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5521 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     5581 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5641 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5701 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     5761 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     5821 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     5881 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     5941 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     6001 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6061 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6121 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6181 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6241 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6301 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6361 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     6421 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6481 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6541 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6601 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6661 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6721 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6781 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6841 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6901 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6961 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     7021 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     7081 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     7141 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7201 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     7261 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7321 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     7381 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     7441 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7501 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7561 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7621 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7681 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7741 ttcaagaact ctgtag
//
