LOCUS       sequence-228220-        7138 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 7138)
  AUTHORS   Gupta PB, Kuperwasser C, Brunet JP, Ramaswamy S, Kuo WL, Gray JW, 
            Naber SP, Weinberg RA
  TITLE     The melanocyte differentiation program predisposes to metastasis 
            after neoplastic transformation.
  JOURNAL   Nat Genet. 2005 Oct . 37(10):1047-54.
  PUBMED    16142232
REFERENCE   2  (bases 1 to 7138)
  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: "Nat Genet. 
            2005 Oct . 37(10):1047-54."
COMMENT     Sequence Label: pBABE-puro TPR-Met
FEATURES             Location/Qualifiers
     source          1..7138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1858..1878)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1863..2192
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2043..2178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2202..2801
                     /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(2202..2221)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2698..2718
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3022..3491
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3621..3638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3792..4380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3872..3891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4540..5331,5332..5400))
                     /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     5163..5182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5401..5505)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5536..6005
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6070..6269
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6470..6886
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6816..6838
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6854..6870
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 atggcggcgg tgttgcagca agtcctggag cgcacggagc tgaacaagct gcccaagtct
       61 gtccagaaca aacttgaaaa gttccttgct gatcagcaat ccgagatcga tggcctgaag
      121 gggcggcatg agaaatttaa ggtggagagc gaacaacagt attttgaaat agaaaagagg
      181 ttgtcccaca gtcaggagag acttgtgaat gaaacccgag agtgtcaaag cttgcggctt
      241 gagctagaga aactcaacaa tcaactgaag gcactaactg agaaaaacaa agaacttgaa
      301 attgctcagg atcgcaatat tgccattcag agccaattta caagaacaaa ggaagaatta
      361 gaagctgaga aaagagactt aattagaacc aatgagagac tatctcaaga acttgaatac
      421 ttaacagatc agtttcctaa ttcatctcag aacggttcat gccgacaagt gcagtatcct
      481 ctgacagaca tgtcccccat cctaactagt ggggactctg atatatccag tccattactg
      541 caaaatactg tccacattga cctcagtgct ctaaatccag agctggtcca ggcagtgcag
      601 catgtagtga ttgggcccag tagcctgatt gtgcatttca atgaagtcat aggaagaggg
      661 cattttggtt gtgtatatca tgggactttg ttggacaatg atggcaagaa aattcactgt
      721 gctgtgaaat ccttgaacag aatcactgac ataggagaag tttcccaatt tctgaccgag
      781 ggaatcatca tgaaagattt tagtcatccc aatgtcctct cgctcctggg aatctgcctg
      841 cgaagtgaag ggtctccgct ggtggtccta ccatacatga aacatggaga tcttcgaaat
      901 ttcattcgaa atgagactca taatccaact gtaaaagatc ttattggctt tggtcttcaa
      961 gtagccaaag gcatgaaata tcttgcaagc aaaaagtttg tccacagaga cttggctgca
     1021 agaaactgta tgctggatga aaaattcaca gtcaaggttg ctgattttgg tcttgccaga
     1081 gacatgtatg ataaagaata ctatagtgta cacaacaaaa caggtgcaaa gctgccagtg
     1141 aagtggatgg ctttggaaag tctgcaaact caaaagttta ccaccaagtc agatgtgtgg
     1201 tcctttggcg tgctcctctg ggagctgatg acaagaggag ccccacctta tcctgacgta
     1261 aacacctttg atataactgt ttacttgttg caagggagaa gactcctaca acccgaatac
     1321 tgcccagacc ccttatatga agtaatgcta aaatgctggc accctaaagc cgaaatgcgc
     1381 ccatcctttt ctgaactggt gtcccggata tcagcgatct tctctacttt cattggggag
     1441 cactatgtcc atgtgaacgc tacttatgtg aacgtaaaat gtgtcgctcc gtatccttct
     1501 ctgttgtcat cagaagataa cgctgatgat gaggtggaca cacgaccagc ctccttctgg
     1561 gagacatcat agtgctagta ctatgtcaaa gcaacagtcc acactttgtc caatggtttt
     1621 ttcactgcct gacctttaaa aggccatcga tattctttgc tcttgccaaa attgcactat
     1681 tataggactt gtattgttat ttaaattact ggattctaag gaatttctta tctgacagag
     1741 catcagaacc agaggcttgg tcccacaggc cacggaccaa tggcctgcag ccgtgacaac
     1801 actcctgtca tattggagtc caaaacttga attcgccagc acagtggtcg accctgtgga
     1861 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     1921 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     1981 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2041 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2101 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2161 gaggcttttt tggaggccta ggcttttgca aaaagcttac catgaccgag tacaagccca
     2221 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     2281 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     2341 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     2401 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     2461 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     2521 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     2581 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     2641 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     2701 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     2761 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc
     2821 agcgcccgac cgaaaggagc gcacgacccc atgcatcgat aaaataaaag attttattta
     2881 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     2941 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa
     3001 ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     3061 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat
     3121 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     3181 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     3241 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     3301 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc
     3361 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     3421 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     3481 ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac
     3541 attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3601 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3661 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3721 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3781 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3841 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3901 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3961 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4021 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4081 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4141 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4201 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4261 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4321 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4381 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4441 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccgcaa
     4501 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4561 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4621 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4681 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4741 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4801 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4861 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4921 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4981 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5041 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5101 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5161 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5221 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5281 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5341 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5401 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5461 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5521 agtgccacct gcagcctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     5581 ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     5641 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc
     5701 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     5761 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     5821 ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg
     5881 actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     5941 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     6001 tttcatttgg gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca
     6061 ccgggaggca agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac
     6121 tgattttatg cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg
     6181 tggtggaact gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt
     6241 tgggggccgt ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc
     6301 aggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt
     6361 tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt
     6421 tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt
     6481 taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa
     6541 ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac
     6601 ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa
     6661 gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac
     6721 ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc
     6781 tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc
     6841 gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccggcc ggatcccagt
     6901 gtggtggtac gtaggaattc cgggggcggt gaggtgctgt tgctgaaacg ctgccgctga
     6961 gggtggactc gatttcccag ggtcccgccg cgggagtctc cggcgggcgg gcgcgcgcga
     7021 gccaccgagc gaggtgatag aggcggcggc ccaggcgtct gggtcctgct ggtcttcgcc
     7081 tttcttctcc gcttctaccc cgtcggccgc tgccactggg gtccctggcc ccaccgac
//
