LOCUS       sequence-256779-        8870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Forced expression of ASAP2f.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8870)
  AUTHORS   Jiang Y, Bao X, Lian XL
  TITLE     A dual cardiomyocyte reporter model derived from human pluripotent 
            stem cells.
  JOURNAL   Stem Cell Res Ther. 2021 May 29;12(1):305. doi: 
            10.1186/s13287-021-02341-6.
  PUBMED    34051863
REFERENCE   2  (bases 1 to 8870)
  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: "Stem Cell 
            Res Ther."; date: "2021-05-29"; volume: "12(1):305. doi"; pages: " 
            10.1186/s13287-021-02341-6"
COMMENT     Sequence Label: pYJ21-lenti-EF1a-ASAP2f-puro
FEATURES             Location/Qualifiers
     source          1..8870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             117..750
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    797..922
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1415..1648
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1833..1877
                     /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    2164..2281
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2410..3588
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2641..3579
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3536..3556
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    join(4986..5560,5561..5563,5564..5572)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5153..5170)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5380..5399
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5592..6191
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5592..5611)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6088..6108
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6394..6627
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6778..6795)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6784..7008
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      7051..7060
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        7104..7208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7209..7277,7278..8069)
                     /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(7427..7446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8240..8828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8729..8748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggaccgcca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
       61 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttgga
      121 agggctaatt cactcccaaa gaagacaaga tatccttgat ctgtggatct accacacaca
      181 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac
      241 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa
      301 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
      361 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct
      421 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg
      481 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg
      541 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
      601 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      661 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      721 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
      781 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
      841 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
      901 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
      961 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1021 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1081 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1141 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1201 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1261 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     1321 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     1381 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     1441 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     1501 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     1561 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     1621 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     1681 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     1741 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     1801 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     1861 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     1921 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     1981 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     2041 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     2101 gatccattcg attagtgaac ggatctcgac ggtatcgcca caaatggcag tattcatcca
     2161 caattttaaa agaaaggggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     2221 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     2281 tcgggtttat tacagggaca gcagagatcc actttggatc gataagcttt gcaaagatgg
     2341 ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga
     2401 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     2461 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
     2521 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     2581 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     2641 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
     2701 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
     2761 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
     2821 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
     2881 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
     2941 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
     3001 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
     3061 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
     3121 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
     3181 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
     3241 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
     3301 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
     3361 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
     3421 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
     3481 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
     3541 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg aattgatcct
     3601 tcgaactaga tggagacgac tgtgaggtat gaacaggggt cagagctcac taaaacttcg
     3661 agctctccaa cagcagatga gcccacgata aagattgatg atggtcgtga tgagggtaat
     3721 gaacaagaca gctgttccaa taccattagg agaaaaattt ccccgtttgt gatgtcattt
     3781 ggattcagag tatttggagt tgtgcttatc attgtagaca tcatagtggt gattgtggat
     3841 ctggccatca gtgagaagaa aagaggcatt agagagattc ttgaaggtgt ttccctggct
     3901 atagcactct tcttccttgt tgatgttctc atgagagtgt ttgttgaagg cttcaagaac
     3961 tatttccggt ccaaactgaa tactttggat gcagtcatag tagtgggcac tctgctaatt
     4021 aatatgacca actccttctc tgaccttagc tttaacagcc ataacgtgta tattaccgcg
     4081 gataaacaga aaaacggcat taaagcgaac tttaccgtgc gccataacgt ggaagatggc
     4141 agcgtgcagc tggcggatca ttatcagcag aacaccccga ttggcgatgg cccggtgctg
     4201 ctgccggata accattatct gagcacccag accgtgctga gcaaagatcc gaacgaaaaa
     4261 cgcgatcaca tggtgctgct ggaatttgtg accgcagcgg gcattacaca cggcatggat
     4321 gaactgtatg gcggcaccgg cggcagcgcg agccagggcg aagaactgtt taccggcgtg
     4381 gtgccgattc tggtggaact ggatggcgat gtgaacggcc ataaatttag cgtgcgcggc
     4441 gaaggcgaag gcgatgcgac cattggcaaa ctgaccctga aatttatttg caccaccggc
     4501 aaactaccgg tgccgtggcc gaccctggtg accaccttaa cctatggcgt gcagtgcttt
     4561 agccgctatc cggatcatat gaaacgccat gattttttta aaagcgcgat gccggaaggc
     4621 tatgtgcagg aacgcaccat tagctttaaa gatgatggca aatataaaac ccgcgcggtg
     4681 gtgaaatttg aaggcgatac cctggtgaac cgcattgaac tgaaaggcac cgattttaaa
     4741 gaagatggca acattctggg gcataaactg gaatataaca cagatcagat gccgcagatg
     4801 gttactcttc ttcgagttct gagaattgtt atcttaataa gaatatttcg cctggcttca
     4861 cagaagaaac aacttgaagt ggtaacctaa aattcgaagc ttgagctcga gatctgagtc
     4921 cggtagcgct agttcgaagg atccgcatgc atctagggcg gccaattccg cccctctccc
     4981 ccccccccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     5041 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     5101 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     5161 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     5221 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     5281 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     5341 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     5401 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     5461 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     5521 cacggggacg tggttttcct ttgaaaaaca cgatgataag cttgccacaa cccacaagga
     5581 gacgaccttc catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     5641 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     5701 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     5761 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     5821 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     5881 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     5941 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     6001 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     6061 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     6121 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     6181 ccggtgcctg ataataggcg gccgctcgag acctagaaaa acatggagca atcacaagta
     6241 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
     6301 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
     6361 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     6421 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6481 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6541 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6601 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6661 tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattat aatagattta
     6721 gcaggaattg aactaggagt ggagcacaca ggcaaagttc tagagctcgc tgatcagcct
     6781 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     6841 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     6901 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     6961 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     7021 aaagaaccag ctgggggcgc gcccctcgag gccgccatgg tcatagctgt ttgacgtcag
     7081 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     7141 caaatatgta tccgctcacg agacaataac cctgataaat gcttcaataa tattgaaaaa
     7201 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     7261 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     7321 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     7381 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     7441 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     7501 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     7561 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7621 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7681 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7741 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7801 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7861 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7921 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7981 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     8041 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     8101 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     8161 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     8221 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     8281 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     8341 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     8401 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     8461 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     8521 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     8581 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8641 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8701 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8761 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8821 tatggaaaaa cgccagcaac gcggcctttt tacggttctg gccttttgca
//
