LOCUS       sequence-140119-        7576 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 7576)
  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 7576)
  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: LPCX Smad2
FEATURES             Location/Qualifiers
     source          1..7576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             414..589
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    652..851
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1052..1468
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1398..1420
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1436..1452
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1494..2093
                     /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(1494..1513)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1990..2010
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     enhancer        2285..2664
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2665..2868
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2819..2839
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2865..2889
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2991..3014
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             4478..5071
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5174..5196)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5282..5422
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5337..5356
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5437..5454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5608..6196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5688..5707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6367..7158,7159..7227))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6990..7009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7228..7332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7400..7418
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1441 tccagccctc actccttctc taggcgccgg aattccgatc tgatcagctt accatgaccg
     1501 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     1561 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     1621 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     1681 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     1741 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     1801 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     1861 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     1921 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     1981 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     2041 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
     2101 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg gatccggcca
     2161 ttagccatat tattcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     2221 acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac attaccgcca
     2281 tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     2341 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     2401 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2461 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     2521 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     2581 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     2641 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2701 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2761 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2821 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2881 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     2941 cgatccagcc tccgcggccc caagccctcg agggcttgtt cgataccatg gactacaagg
     3001 acgacgatga caagggtcga caatcgtcca tcttgccatt cacgccgcca gttgtgaaga
     3061 gactgctggg atggaagaag tcagctggtg ggtctggagg agcaggcgga ggagagcaga
     3121 atgggcagga agaaaagtgg tgtgagaaag cagtgaaaag tctggtgaag aagctaaaga
     3181 aaacaggacg attagatgag cttgagaaag ccatcaccac tcaaaactgt aatactaaat
     3241 gtgttaccat accaagcact tgctctgaaa tttggggact gagtacacca aatacgatag
     3301 atcagtggga tacaacaggc ctttacagct tctctgaaca aaccaggtct cttgatggtc
     3361 gtctccaggt atcccatcga aaaggattgc cacatgttat atattgccga ttatggcgct
     3421 ggcctgatct tcacagtcat catgaactca aggcaattga aaactgcgaa tatgctttta
     3481 atcttaaaaa ggatgaagta tgtgtaaacc cttaccacta tcagagagtt gagacaccag
     3541 ttttgcctcc agtattagtg ccccgacaca ccgagatcct aacagaactt ccgcctctgg
     3601 atgactatac tcactccatt ccagaaaaca ctaacttccc agcaggaatt gagccacaga
     3661 gtaattatat tccagaaacg ccacctcctg gatatatcag tgaagatgga gaaacaagtg
     3721 accaacagtt gaatcaaagt atggacacag gctctccagc agaactatct cctactactc
     3781 tttcccctgt taatcatagc ttggatttac agccagttac ttactcagaa cctgcatttt
     3841 ggtgttcgat agcatattat gaattaaatc agagggttgg agaaaccttc catgcatcac
     3901 agccctcact cactgtagat ggctttacag acccatcaaa ttcagagagg ttctgcttag
     3961 gtttactctc caatgttaac cgaaatgcca cggtagaaat gacaagaagg catataggaa
     4021 gaggagtgcg cttatactac ataggtgggg aagtttttgc tgagtgccta agtgatagtg
     4081 caatctttgt gcagagcccc aattgtaatc agagatatgg ctggcaccct gcaacagtgt
     4141 gtaaaattcc accaggctgt aatctgaaga tcttcaacaa ccaggaattt gctgctcttc
     4201 tggctcagtc tgttaatcag ggttttgaag ccgtctatca gctaactaga atgtgcacca
     4261 taagaatgag ttttgtgaaa gggtggggag cagaataccg aaggcagacg gtaacaagta
     4321 ctccttgctg gattgaactt catctgaatg gacctctaca gtggttggac aaagtattaa
     4381 ctcagatggg atccccttca gtgcgttgct caagcatgtc ataatctaga acatcgataa
     4441 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     4501 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa
     4561 tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga
     4621 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat
     4681 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     4741 atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag
     4801 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     4861 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     4921 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     4981 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     5041 attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc gggagacccc
     5101 tgcccaggga ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg cgtttcggtg
     5161 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     5221 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     5281 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     5341 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     5401 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5461 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5521 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5581 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5641 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5701 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5761 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5821 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5881 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5941 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6001 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6061 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6121 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttactcgcag
     6181 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6241 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6301 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6361 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6421 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6481 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6541 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6601 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6661 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6721 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6781 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6841 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6901 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6961 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     7021 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7081 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7141 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7201 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7261 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7321 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     7381 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattcat
     7441 accagatcac cgaaaactgt cctccaaatg tgtccccctc acactcccaa attcgcgggc
     7501 ttctgcctct tagaccactc taccctattc cccacactca ccggagccaa agccgcggcc
     7561 cttccgtttc tttgct
//
