LOCUS       sequence-402627-        6462 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 6462)
  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 6462)
  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 Smad7
FEATURES             Location/Qualifiers
     source          1..6462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     306..326
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(1712..1735)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1758..2174)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(1774..1790)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1806..1828)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    2375..2574
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             2639..3108
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        3139..3243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3244..3312,3313..4104)
                     /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(3462..3481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4264..4852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4753..4772
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5006..5023
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             5153..5622
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(5845..6444)
                     /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(5928..5948)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6425..6444
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(6454..6462,1..321))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
       61 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
      121 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      181 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      241 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      301 tccacaccct aactgacaca cattccacag ggtcgacccc atcgatggga gatctcccga
      361 attagcttct gcaggtcgag tcacgatccg ccctgtcctc ttctccccac cacacgaccg
      421 actaccggct gttgaagatg acctccagcc agcacgggca gctgctgatg aactggcggg
      481 tgtagcactg gccccagccc ttcacaaagc tgatctgcac ggtgaaaccc gtccatggtt
      541 gctgcatgaa ctcgtggtca ttgggccgct gcaggctgta ggctttctca tagtcaaaag
      601 ccttgatgga gaaaccaggg aacactttgt gcaccaacag cgtcctggag tccgggttgt
      661 ccagtgtggc ggacttgatg aagatggggt aactgctgcg gttgtaaacc cacacgccat
      721 ccacttcccg cgtcagctgg atgccacagc cgatcttgct ccgcactttc tgtaccagct
      781 gactcttgtt gtccgaattg agctgtccga ggcaaaagcc attcccctga ggtagatcat
      841 agaagatatc cagggagggc tcttggacac agtagagcct ccccacgcga gtcttctcct
      901 cccagtatgc caccacgcac cagtgtgacc gatccccagg ctccagaaga agttgggaat
      961 ctgaaagccc cccaggggcc agataattcg ttcccccggt ttccgcggag gaaggtacag
     1021 catctggaca gcctgcagtt ggtttgagaa aatccattgg gtatctggag taaggaggag
     1081 ggggagactc tagttcacag agtcgactaa ggtgatgggg gttgcagcac accagctcgg
     1141 ggttgatctt cccgtaagat tcacagcaac acagcctctt gacttccgag gaatgcctga
     1201 gatccggcca cctgaacact ttgcacagca ggagggggag cgagtaggac gagggcggct
     1261 gcgcgggctg cgcgctggcg ggcgcccccg ggcccagcct gcagtccagg cggccgggca
     1321 gcaggaggca cgcggtgcgc gtaccgccgc gggactccac ggcctgaagc agcagctcca
     1381 gctgccgctc cttgagtttc ttgagcaccg agtgcgtgag cgccttcaga tccgcctcgg
     1441 cgcccccggc cgccccggca cccgaggttg ggggatgggg atggtggtga cctttggcac
     1501 ctcggactgc cttgcccagg cagcagccag ccctgcccgc accgccgcca ccagccccat
     1561 aagcccggcc gtccgtcgcc ccttctcccc gcagctcgcc tcctccgccg ccacccccca
     1621 cgccctcctc ctcgtcctcg ccgccgggcg cacggctcct ccagagacgc cggacgagcg
     1681 cagatcgttt ggtcctgaac ccgaattcgc ccttgtcatc atcgtccttg tagtccatgg
     1741 cggtggatca tccggccggc gcctagagaa ggagtgaggg ctggataaag ggaggatcga
     1801 ggcggggtcg aacgaggagg ttcaaggggg agagacgggg cggatggagg aagaggaggc
     1861 ggaggcttag ggtgtacaaa gggcttgacc cagggagggg ggtcaaaagc caaggcttcc
     1921 caggtcacga tgtaggggac ctggtctggg tgtccatgcg ggccaggtga aaagaccttg
     1981 atcttaacct gggtgatgag gtctcggtta aaggtgccgt ctcgcggcca tccgacgtta
     2041 aaggttggcc attctgcaga gcagaaggta acccaacgtc tcttcttgac atctaccgac
     2101 tggttgtgag cgatccgctc gacatctttc cagtgaccta aggtcaaact taagggagtg
     2161 gtaacagtct ggccctaatt ttcagacaaa tacagaaaca cagtcagaca gagacaacac
     2221 agaacgatgc tgcagcgctg cagcagacaa gacgcgcggc ttcggttcca aaccgaaagc
     2281 aaaaattcag acggaggcgg gaactgtttt aggttctcgt ctcctaccag aaccacatat
     2341 cctgacgggg tcggattcca catcgactcc cttcctcagg tcgggccaca aaaacggccc
     2401 ccaaagtccc tgggacgtct cccagggttg cggccgggtg ttcagaactc gtcagttcca
     2461 ccacgggtcc gccagataca gagctagtta gctaactagt accgacgcag gcgcataaaa
     2521 tcagtcatag acactagaca atcggacaga cacagataag ttgctggcca gcttgcctcc
     2581 cggtggtggg tcggtggtcc ctgggcaggg gtctcccgat cccggacgag cccccaaatg
     2641 aaagaccccc gctgacgggt agtcaatcac tcagaggaga ccctcccaag gaacagcgag
     2701 accacaagtc ggatgcaact gcaagagggt ttattggata cacgggtacc cgggcgactc
     2761 agtcaatcgg aggactggcg ccccgagtga ggggttgtgg gctcttttat tgagctcggg
     2821 gagcagaagc gcgcgaacag aagcgagaag cgaactgatt ggttagttca aataaggcac
     2881 agggtcattt caggtccttg gggcaccctg gaaacatctg atggttctct agaaactgct
     2941 gagggctgga ccgcatctgg ggaccatctg ttcttggccc tgagccgggg caggaactgc
     3001 ttaccacaga tatcctgttt ggcccatatt cagctgttcc atctgttctt ggccctgagc
     3061 cggggcagga actgcttacc acagatatcc tgtttggccc atattcaggc tgcaggtggc
     3121 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3181 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3241 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3301 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3361 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3421 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3481 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3541 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3601 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3661 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3721 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3781 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3841 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3901 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3961 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4021 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4081 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4141 gatttgcggc cgcaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4201 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4261 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4321 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4381 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4441 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4501 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4561 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4621 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4681 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4741 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4801 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4861 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4921 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4981 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     5041 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcaact atggccattt
     5101 aatgtaaata cttaagaaaa aaaaccaaat taattttgat acatgctgca tgtgaaagac
     5161 ccccgctgac gggtagtcaa tcactcagag gagaccctcc caaggaacag cgagaccaca
     5221 agtcggatgc aactgcaaga gggtttattg gatacacggg tacccgggcg actcagtcaa
     5281 tcggaggact ggcgccccga gtgaggggtt gtgggctctt ttattgagct cggggagcag
     5341 aagcgcgcga acagaagcga gaagcgaact gattggttag ttcaaataag gcacagggtc
     5401 atttcaggtc cttggggcac cctggaaaca tctgatggtt ctctagaaac tgctgagggc
     5461 tggaccgcat ctggggacca tctgttcttg gccctgagcc ggggcaggaa ctgcttacca
     5521 cagatatcct gtttggccca tattcagctg ttccatctgt tcttggccct gagccggggc
     5581 aggaactgct taccacagat atcctgtttg gcccatattc agctgttcca tctgttcctg
     5641 accttgatct gaacttctct attctcagtt atgtattttt ccatgccttg caaaatggcg
     5701 ttacttaagc tagcttgcca aacctacagg tggggtcttt cattcccccc tttttctgga
     5761 gactaaataa aatcttttat tttatcgcga tgcatggggt cgtgcgctcc tttcggtcgg
     5821 gcgctgcggg tcgtggggcg ggcgtcaggc accgggcttg cgggtcatgc accaggtgcg
     5881 cggtccttcg ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg
     5941 gaggttgcgg ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc
     6001 cactccgggg agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc
     6061 gacggtggcc aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat
     6121 ctgttgctgc gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc
     6181 gaacaccgcc cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc
     6241 gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag
     6301 ctcggtgacc cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg cggggtagtc
     6361 ggcgaacgcg gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg
     6421 caccgtgggc ttgtactcgg tcatggtaag ctttttgcaa aa
//
