LOCUS       sequence-456517-        7319 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 7319)
  AUTHORS   Yang Z, MacQuarrie KL, Analau E, Tyler AE, Dilworth FJ, Cao Y, Diede
            SJ, Tapscott SJ
  TITLE     MyoD and E-protein heterodimers switch rhabdomyosarcoma cells from 
            an arrested myoblast phase to a differentiated state.
  JOURNAL   Genes Dev. 2009 Mar 15. 23(6):694-707.
  PUBMED    19299559
REFERENCE   2  (bases 1 to 7319)
  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: "Genes Dev. 
            2009 Mar 15. 23(6):694-707."
COMMENT     Sequence Label: E12-pCLBabe
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        378..581
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     532..552
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    820..1019
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1220..1636
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1566..1588
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1604..1620
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3623..3643)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3628..3957
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3808..3943
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3967..4566
                     /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(3967..3986)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4463..4483
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4787..5256
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5386..5403)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5557..6145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5637..5656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6305..7096,7097..7165))
                     /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     6928..6947
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        join(7317..7319,1..377)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
       61 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      121 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      181 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      241 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      301 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      361 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      421 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      481 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      541 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcaataaaag agcccacaac
      601 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
      661 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
      721 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
      781 gacccctgcc cagggaccac cgacccacca ccgggaggca agctggccag caacttatct
      841 gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt
      901 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct gaacacccgg
      961 ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc cgacctgagg
     1021 aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag gagacgagaa
     1081 cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc
     1141 gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct
     1201 gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca
     1261 ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg
     1321 ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac
     1381 ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg
     1441 acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc
     1501 ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc
     1561 tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac
     1621 tccttctcta ggcgccggcc ggatccatgg cgcctgtggg cacagacaag gagctcagtg
     1681 acctcctgga cttcagcatg atgttcccgc tgcctgtcac caacgggaag ggccggcccg
     1741 cctccctggc cggggcgcag ttcggaggtt caggtcttga ggaccggccc agctcaggct
     1801 cctggggcag cggcgaccag agcagctcct cctttgaccc cagccggacc ttcagcgagg
     1861 gcacccactt cactgagtcg cacagcagcc tctcttcatc cacattcctg ggaccgggac
     1921 tcggaggcaa gagcggtgag cggggcgcct atgcctcctt cgggagagac gcaggcgtgg
     1981 gcggcctgac tcaggctggc ttcctgtcag gcgagctggc cctcaacagc cccgggcccc
     2041 tgtccccttc gggcatgaag gggacctccc agtactaccc ctcctactcc gacagctccc
     2101 ggcggagagc ggcagacggc agcctagaca cgcagcccaa gaaggtccgg aaggtcccgc
     2161 cgggtcttcc atcctcggtg tacccaccca gctcaggtga ggactacggc agggatgcca
     2221 ccgcctaccc gtccgccaag acccccagca gcacctatcc cgcccccttc tacgtggcag
     2281 atggcagcct gcacccctca gccgagctct ggagtccccc gggccaggcg ggcttcgggc
     2341 ccatgctggg tgggggctca tccccgctgc ccctcccgcc cggtagcggc ccggtgggca
     2401 gcagtggaag cagcagcacg tttggtggcc tgcaccagca cgagcgtatg ggctaccagc
     2461 tgcatggagc agaggtgaac ggtgggctcc catctgcatc ctccttctcc tcagcccccg
     2521 gagccacgta cggcggcgtc tccagccaca cgccgcctgt cagcggggcc gacagcctcc
     2581 tgggctcccg agggaccaca gctggcagct ccggggatgc cctcggcaaa gcactggcct
     2641 cgatctactc cccggatcac tcaagcaata acttctcgtc cagcccttct acccccgtgg
     2701 gctcccccca gggcctggca ggaacgtcac agtggcctcg agcaggagcc cccggtgcct
     2761 tatcgcccag ctacgacggg ggtctccacg gcctgcagag taagatagaa gaccacctgg
     2821 acgaggccat ccacgtgctc cgcagccacg ccgtgggcac agccggcgac atgcacacgc
     2881 tgctgcctgg ccacggggcg ctggcctcag gtttcaccgg ccccatgtca ctgggcgggc
     2941 ggcacgcagg cctggttgga ggcagccacc ccgaggacgg cctcgcaggc agcaccagcc
     3001 tcatgcacaa ccacgcggcc ctccccagcc agccaggcac cctccctgac ctgtctcggc
     3061 ctcccgactc ctacagtggg ctagggcgag caggtgccac ggcggccgcc agcgagatca
     3121 agcgggagga gaaggaggac gaggagaaca cgtcagcggc tgaccactcg gaggaggaga
     3181 agaaggagct gaaggccccc cgggcccgga ccagcccaga cgaggacgag gacgaccttc
     3241 tccccccaga gcagaaggcc gagcgggaga aggagcgccg ggtggccaat aacgcccggg
     3301 agcggctgcg ggtccgtgac atcaacgagg cctttaagga gctggggcgc atgtgccaac
     3361 tgcacctcaa cagcgagaag ccccagacca aactgctcat cctgcaccag gctgtctcgg
     3421 tcatcctgaa cttggagcag caagtgcgag agcggaacct gaatcccaaa gcagcctgtt
     3481 tgaaacggcg agaagaggaa aaggtgtcag gtgtggttgg agacccccag atggtgcttt
     3541 cagctcccca cccaggcctg agcgaagccc acaaccccgc cgggcacatg tgagaattcg
     3601 ccagcacagt ggtcgaccct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3661 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3721 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3781 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3841 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3901 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3961 cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     4021 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     4081 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     4141 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     4201 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     4261 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     4321 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     4381 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     4441 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4501 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     4561 gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca
     4621 tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     4681 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     4741 ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca
     4801 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca
     4861 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     4921 agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg
     4981 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     5041 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     5101 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
     5161 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     5221 ctgagtgatt gactacccgt cagcgggggt ctttcacatg cagcatgtat caaaattaat
     5281 ttggtttttt ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca
     5341 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5401 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5461 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5521 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5581 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5641 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5701 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5761 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5821 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5881 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5941 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6001 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6061 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6121 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6181 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6241 cacctagatc cttttgcggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg
     6301 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6361 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6421 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6481 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6541 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6601 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6661 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6721 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6781 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6841 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6901 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6961 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7021 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7081 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7141 cgacacggaa atgttgaata ctcatactct tcctttttca atgatccggc cattagccat
     7201 attattcatt ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta
     7261 tccatatcat aatatgtaca tttatattgg ctcatgtcca acattaccgc catgttgac
//
