LOCUS       Exported               10055 bp ds-DNA     circular SYN 16-AUG-2021
DEFINITION  Retroviral expression vector for murine direct cardiac 
            reprogramming.
ACCESSION   .
VERSION     .
KEYWORDS    pmx-puro-MiGT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10055)
  AUTHORS   Wang L, Huang P, Near D, Ravi K, Xu Y, Liu J, Qian L
  TITLE     Isoform Specific Effects of Mef2C during Direct Cardiac 
            Reprogramming.
  JOURNAL   Cells. 2020 Jan 22;9(2). pii: cells9020268. doi: 
            10.3390/cells9020268.
  PUBMED    31979018
REFERENCE   2  (bases 1 to 10055)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 16, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10055
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1832..1848
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             3339..3395
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4740..4793
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(6390..6410)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6395..6724
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6575..6710
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6637..6656
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6734..7333
                     /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(6734..6753)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7230..7250
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             7560..8029
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(8159..8176)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8330..8918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8410..8429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9082..9942)
                     /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     9705..9724
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9943..10047)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
     1921 ccgccatggg gagaaaaaag attcagatta cgaggataat ggatgagcgt aacagacagg
     1981 tgacttttac gaagaggaaa tttggattga tgaagaaggc ttatgagctg agcgtgctgt
     2041 gtgactgtga gattgcactg atcatcttca acagcaccaa caagctgttc cagtacgcca
     2101 gcactgacat ggataaggtg ttgctcaagt acaccgagta caacgagccg cacgagagcc
     2161 ggacaaactc agacattgtg gagacattga gaaagaaggg cctcaatggc tgtgacagcc
     2221 cagatcccga tgcagacgat tcagtaggtc acagccctga gtctgaggac aagtacagga
     2281 aaattaacga agatattgat ctaatgatca gcaggcaaag attgtgtgct gttccacctc
     2341 ccagctttga gatgccagtt accatcccag tgtccagcca taacagtttg gtgtacagca
     2401 atcctgtcag cacactggga aaccccaatc ttctgccact ggcccacccg tctctgcaga
     2461 ggaatagtat gtctcctggt gtaacacata gacctccaag tgcaggtaac acaggcggtc
     2521 tgatgggcgg agatctgaca tccggtgcag gcaccagcgc agggaatgga tacggcaacc
     2581 cccggaactc accaggcctg ctggtctcac ctggtaacct gaacaagaat atacaagcca
     2641 aatctcctcc ccctatgaat ctaggaatga ataatcgtaa gccagatctc cgcgttctta
     2701 tcccacctgg cagcaagaac acgatgccat cagtgaatca aaggataaat aactcccagt
     2761 cggctcagtc attggctacc ccggtggttt ccgtagcaac tcctacttta ccaggacaag
     2821 gaatgggagg atatccatca gccatttcaa caacatatgg tactgagtac tctctgagta
     2881 gcgcagatct gtcatctctg tctggcttca acactgccag tgcgctccac ctcggctctg
     2941 taactggctg gcagcagcag cacctacata acatgccgcc atctgccctc agtcagttgg
     3001 gagcttgcac tagcactcat ttatctcaga gttcaaatct ctccctgcct tctactcaaa
     3061 gcctcagcat caagtcagaa cctgtttctc ctcctagaga ccgtaccacc accccttcga
     3121 gatacccaca acacaccacg cgccacgagg cggggaggtc tcctgttgac agcttgagca
     3181 gctgtagcag ttcctacgat gggagcgacc gagaggatca ccggaacgaa ttccactccc
     3241 ccattggact caccagacct tcgccggacg aaagggaaag tccttcagtc aagcgcatgc
     3301 gactctctga aggatgggca acagctagcg gcagcggcgc cacgaacttc tctctgttaa
     3361 agcaagcagg agatgttgaa gaaaaccccg ggcctgcatg catgtaccaa agcctggcca
     3421 tggccgccaa ccacggcccc ccgcccggcg cctacgaagc aggtggccct ggcgccttca
     3481 tgcacagcgc gggcgccgcg tcctcgcccg tctacgtgcc cactccgcgg gtgccgtcct
     3541 ctgtgctggg cctgtcctac ctgcagggcg gtggcagtgc cgctgcagct ggaaccacct
     3601 cgggtggcag ctccggggcc ggcccgtcgg gtgcagggcc tgggacccag cagggtagcc
     3661 ctggctggag ccaagctgga gccgagggag ccgcctacac cccgccgccc gtgtccccgc
     3721 gcttctcttt cccggggact actgggtccc tggcggccgc tgccgccgct gccgcagccc
     3781 gggaagctgc agcctacggc agtggcggcg gggcggcggg cgctggtctg gctggccgag
     3841 agcagtacgg gcgtccgggc ttcgccggct cctactccag cccctaccca gcctacatgg
     3901 ccgacgtggg agcatcctgg gccgcagccg ctgccgcctc tgccggcccc ttcgacagcc
     3961 cagtcctgca cagcctgcct ggacgggcca accctggaag acaccccaat ctcgatatgt
     4021 ttgatgactt ctcagaaggc agagagtgtg tcaattgtgg ggccatgtcc accccactct
     4081 ggaggcgaga tgggacggga cactacctgt gcaatgcctg tggcctctat cacaagatga
     4141 acggcatcaa ccggcccctc attaagcctc agcgccgcct gtccgcttcc cgccgggtag
     4201 gcctctcctg tgccaactgc cagactacca ccaccacgct gtggcgtcgt aatgccgagg
     4261 gtgagcctgt atgtaatgcc tgcggcctct acatgaagct ccatggggtt cccaggcctc
     4321 ttgcaatgcg gaaggagggg attcaaacca gaaaacggaa gcccaagaac ctgaataaat
     4381 ctaagacgcc agcaggtcct gctggtgaga ccctccctcc ctccagtggt gcctccagcg
     4441 gtaactccag caatgccact agcagcagca gcagcagtga agagatgcgc cccatcaaga
     4501 cagagcccgg gctgtcatct cactatgggc acagcagctc catgtcccag acattcagta
     4561 ctgtgtccgg ccacgggccc tccatccatc cagtgctgtc tgctctgaag ctgtccccac
     4621 aaggctatgc atctcctgtc actcagacat cgcaggccag ctccaagcag gactcttgga
     4681 acagcctggt cctggctgac agtcatgggg acataatcac cgcggcatgc ggctccggcg
     4741 agggcagggg aagtcttcta acatgcgggg acgtggagga aaatcccggc ccactcgaga
     4801 tggccgatac agatgagggc tttggcctgg cgcgcacgcc tctggagcct gattccaaag
     4861 acaggtcttg cgattcgaaa cctgagagtg ctctgggggc tcccagcaag tctccatcat
     4921 ccccgcaggc tgccttcacc cagcagggca tggaaggaat caaggtgttt cttcatgaac
     4981 gtgaactgtg gctgaagttc cacgaagtgg gcacagagat gatcatcacc aaggcaggga
     5041 ggagaatgtt tcctagttac aaagtgaagg tgactggcct taatcccaaa acgaagtata
     5101 ttcttctcat ggatattgtt cccgcagacg accacagata taaatttgct gataacaaat
     5161 ggtccgtaac tggcaaagca gagcctgcca tgccggggcg cctttacgtg cacccggact
     5221 ccccagcaac cggagcccac tggatgcgac aacttgtctc cttccagaag ctcaaactca
     5281 ccaacaacca cctggacccg tttggacaca ttatcctgaa ctccatgcac aaataccagc
     5341 cccgattaca catcgtgaaa gcagacgaaa ataatgggtt cggttcaaag aacactgcgt
     5401 tttgcaccca cgtcttcccg gagacagctt ttatcgctgt gacttcgtac cagaatcaca
     5461 agatcacaca gctgaaaatt gagaacaacc ccttcgccaa aggctttcgg ggcagtgatg
     5521 acctggagtt acacaggatg tctcggatgc aaagtaaaga gtatcctgtg gttcccagga
     5581 gcacagtgag gcacaaagtc acctccaacc acagcccctt cagcagcgag acccgagctc
     5641 tctccacctc atccaattta gggtcccagt accagtgtga gaatggtgtc tctggcccct
     5701 cccaggacct tctgccccca cctaacccat acccactggc ccaggagcac agccaaattt
     5761 accactgtac caagaggaaa gatgaggaat gttccagcac ggagcacccc tataagaagc
     5821 cgtacatgga gacatccccc agcgaggaag acaccttcta tcgctcgggc tacccccagc
     5881 agcagggcct gagtacctct tacaggacag agtcggccca gcggcaggcc tgcatgtatg
     5941 ccagctccgc tccccccagc gagcccgtgc ctagcctgga ggacatcagc tgtaacacat
     6001 ggcccagcat gccctcctat agcagctgta ccgtcaccac cgtgcagccc atggaccgtc
     6061 ttccctacca gcacttctcc gctcatttca cctcggggcc cctggtccct cggttggctg
     6121 gcatggccaa ccatggttct ccccagctcg gcgaagggat gtttcagcac cagacctcag
     6181 tggcccatca gcctgtggtc aggcagtgcg ggcctcagac tggccttcag tctccgggcg
     6241 gcctccagcc cccagagttt ctctacactc acggcgtgcc caggaccctg tccccccatc
     6301 agtatcactc ggtacacggc gtcggcatgg tgccagagtg gagtgagaat agctaagtcg
     6361 agcggccgcc agcacagtgg tcgacctgtg gaatgtgtgt cagttagggt gtggaaagtc
     6421 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     6481 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6541 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6601 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6661 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6721 caaaaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     6781 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     6841 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     6901 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     6961 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     7021 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     7081 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     7141 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     7201 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     7261 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     7321 gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc
     7381 ccatgcatcg tcgacgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     7441 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     7501 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     7561 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     7621 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     7681 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     7741 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     7801 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     7861 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     7921 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     7981 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     8041 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     8101 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     8161 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     8221 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     8281 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     8341 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     8401 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     8461 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     8521 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     8581 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     8641 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     8701 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     8761 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccaggtacc ttcggaaaaa
     8821 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     8881 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8941 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     9001 caaaaaggat cttcacctag atccttttgc ggccggccgc aaatcaatct aaagtatata
     9061 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     9121 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     9181 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     9241 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     9301 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     9361 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     9421 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     9481 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     9541 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     9601 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     9661 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9721 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     9781 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9841 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9901 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9961 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
    10021 gaaaaataaa caaatagggg ttccgcgcac atttc
//
