LOCUS       sequence-146308-        6712 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 6712)
  AUTHORS   Choy L, Skillington J, Derynck R
  TITLE     Roles of autocrine TGF-beta receptor and Smad signaling in adipocyte
            differentiation.
  JOURNAL   J Cell Biol. 2000 May 1. 149(3):667-82.
  PUBMED    10791980
REFERENCE   2  (bases 1 to 6712)
  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: "J Cell 
            Biol. 2000 May 1. 149(3):667-82."
COMMENT     Sequence Label: pBabe Flag Smad6
FEATURES             Location/Qualifiers
     source          1..6712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1378..1401
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2951..2971)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2956..3285
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3136..3271
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3295..3894
                     /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(3295..3314)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3791..3811
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3993..4586
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4787..4804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4958..5546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5038..5057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5724..6515,6516..6584))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6347..6366
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6585..6689)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccac cgccatggac
     1381 tacaaggacg atgatgacaa gggcgaattc gggttcaggt ctaaacgttc ggggctggtg
     1441 cgacgacttt ggcgaagtcg tgtggtccct gatcgggagg aaggcagcgg cggcggcggt
     1501 ggtgtcgacg aggatgggag cctgggcagc cgagctgagc ctgccccgcg ggcacgagag
     1561 ggcggaggct gcagccgctc cgaagtccgc tcggtagccc cgcggcggcc ccgggacgcg
     1621 gtgggaccgc gaggcgccgc gatcgcgggc aggcgccggc gcacaggggg cctcccgagg
     1681 cccgtgtcgg agtcgggggc cggggctggg ggctccccgc tggatgtggc ggagcctgga
     1741 ggcccaggct ggctgcctga gagtgactgc gagacggtga cctgctgtct cttctccgaa
     1801 cgggacgcag caggcgcgcc ccgggactct ggcgatcccc aagccagaca gtccccggag
     1861 ccggaggagg gcggcgggcc tcggagtcgc gaagcccgct cgcgactgct gcttctggag
     1921 caggagctca agacggtcac gtactcgctg ctcaagaggc tcaaggagcg ttcgctggac
     1981 acgctgttgg aggctgtgga gtcccgaggc ggcgtaccgg gcggctgcgt gctggtgccg
     2041 cgcgccgacc tccgcttggg cggccagccc gcgccaccgc agctgctgct cggccgcctc
     2101 ttccgctggc cagacctgca gcacgcagtg gagctgaaac ccctgtgcgg ctgccacagc
     2161 tttaccgccg ccgccgacgg gcccacggtg tgttgcaacc cctaccactt cagccggctc
     2221 tgcgggccag aatcaccgcc gcccccctat tctcggctgt ctcctcctga ccagtacaag
     2281 ccactggatc tgtccgattc tacattgtct tacactgaaa ccgaggccac caactccctc
     2341 atcactgctc cgggtgaatt ctcagatgcc agcatgtctc cggatgccac caagccgagc
     2401 cactggtgca gcgtggcgta ctgggagcac cggacacgcg tgggccgcct ctatgcggtg
     2461 tacgaccagg ctgtcagcat tttctacgac ctacctcagg gcagcggctt ctgcctgggc
     2521 cagctcaacc tggagcagcg cagtgagtcg gtgcggcgca cgcgcagcaa gatcggtttt
     2581 ggcatactgc tcagcaagga gccagacggc gtgtgggcct acaaccgggg cgagcacccc
     2641 atcttcgtca actccccgac gctggatgcg cccggaggcc gcgccctggt cgtgcgcaag
     2701 gtgccaccgg gttactccat caaggtgttc gactttgagc gctcagggct gctgcagcac
     2761 gcagacgccg ctcacggccc ctacgacccg cacagtgtgc gcatcagctt cgccaagggc
     2821 tggggaccct gctactcgcg acagttcatc acctcctgcc cctgttggct ggagatccta
     2881 ctcaacaacc acagatagct cgagagtgtg gtggtacgta ggaattcgcc agcacagtgg
     2941 tcgaccctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3001 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3061 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3121 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3181 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3241 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taccatgacc
     3301 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     3361 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     3421 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     3481 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     3541 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     3601 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     3661 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     3721 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     3781 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     3841 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     3901 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc gataaaataa
     3961 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     4021 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     4081 aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct
     4141 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg
     4201 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     4261 ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     4321 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     4381 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     4441 ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
     4501 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     4561 ctacccgtca gcgggggtct ttcatttccg acttgtggtc tcgctgcctt gggagggtct
     4621 cctctgagtg attgactacc cgtcagcggg ggtcttcaca tgcagcatgt atcaaaatta
     4681 atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc
     4741 caacgcgcgg ggagaggcgg tttgcgtatt ggcgctcttc cgcttcctcg ctcactgact
     4801 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4861 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4921 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4981 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5041 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5101 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5161 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5221 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5281 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5341 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5401 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5461 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5521 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5581 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5641 tcacctagat ccttttaaat taaaaatgaa gtttgcggcc gcaaatcaat ctaaagtata
     5701 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5761 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5821 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5881 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5941 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6001 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6061 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6121 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6181 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6241 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6301 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     6361 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6421 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6481 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6541 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6601 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6661 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc ac
//
