LOCUS       sequence-265933-        6083 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 6083)
  AUTHORS   Zhang Y, Feng XH, Derynck R
  TITLE     Smad3 and Smad4 cooperate with c-Jun/c-Fos to mediate 
            TGF-beta-induced transcription.
  JOURNAL   Nature. 1998 Aug 27. 394(6696):909-13.
  PUBMED    9732876
REFERENCE   2  (bases 1 to 6083)
  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: "Nature. 
            1998 Aug 27. 394(6696):909-13."
COMMENT     Sequence Label: pRK5F Smad2
FEATURES             Location/Qualifiers
     source          1..6083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             911..934
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2370..2379
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2383..2517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2420..2439)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2474..2493
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2586..2943
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2586..2777
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2609..2629)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2794..2929
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2963..2980)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2963..2979)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2972..2994)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3192..3647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3279..3298)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3489..3510
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3664..3683)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3783..3805
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3843..3861)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3929..4033
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4034..4102,4103..4894)
                     /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(4252..4271)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5065..5653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5554..5573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5807..5824
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5941..5962
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5977..5982,5983..6000,6001..6007)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6015..6031
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6020..6042
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6039..6055
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6039..6055
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgataccatg gactacaagg acgacgatga caagggtcga caatcgtcca tcttgccatt
      961 cacgccgcca gttgtgaaga gactgctggg atggaagaag tcagctggtg ggtctggagg
     1021 agcaggcgga ggagagcaga atgggcagga agaaaagtgg tgtgagaaag cagtgaaaag
     1081 tctggtgaag aagctaaaga aaacaggacg attagatgag cttgagaaag ccatcaccac
     1141 tcaaaactgt aatactaaat gtgttaccat accaagcact tgctctgaaa tttggggact
     1201 gagtacacca aatacgatag atcagtggga tacaacaggc ctttacagct tctctgaaca
     1261 aaccaggtct cttgatggtc gtctccaggt atcccatcga aaaggattgc cacatgttat
     1321 atattgccga ttatggcgct ggcctgatct tcacagtcat catgaactca aggcaattga
     1381 aaactgcgaa tatgctttta atcttaaaaa ggatgaagta tgtgtaaacc cttaccacta
     1441 tcagagagtt gagacaccag ttttgcctcc agtattagtg ccccgacaca ccgagatcct
     1501 aacagaactt ccgcctctgg atgactatac tcactccatt ccagaaaaca ctaacttccc
     1561 agcaggaatt gagccacaga gtaattatat tccagaaacg ccacctcctg gatatatcag
     1621 tgaagatgga gaaacaagtg accaacagtt gaatcaaagt atggacacag gctctccagc
     1681 agaactatct cctactactc tttcccctgt taatcatagc ttggatttac agccagttac
     1741 ttactcagaa cctgcatttt ggtgttcgat agcatattat gaattaaatc agagggttgg
     1801 agaaaccttc catgcatcac agccctcact cactgtagat ggctttacag acccatcaaa
     1861 ttcagagagg ttctgcttag gtttactctc caatgttaac cgaaatgcca cggtagaaat
     1921 gacaagaagg catataggaa gaggagtgcg cttatactac ataggtgggg aagtttttgc
     1981 tgagtgccta agtgatagtg caatctttgt gcagagcccc aattgtaatc agagatatgg
     2041 ctggcaccct gcaacagtgt gtaaaattcc accaggctgt aatctgaaga tcttcaacaa
     2101 ccaggaattt gctgctcttc tggctcagtc tgttaatcag ggttttgaag ccgtctatca
     2161 gctaactaga atgtgcacca taagaatgag ttttgtgaaa gggtggggag cagaataccg
     2221 aaggcagacg gtaacaagta ctccttgctg gattgaactt catctgaatg gacctctaca
     2281 gtggttggac aaagtattaa ctcagatggg atccccttca gtgcgttgct caagcatgtc
     2341 ataatctaga gtcgacctgc agaagcttgg ccgccatggc ccaacttgtt tattgcagct
     2401 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2461 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcggg
     2521 aattaattcg gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     2581 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2641 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2701 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2761 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2821 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2881 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2941 aaaaagctgt taacagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     3001 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     3061 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     3121 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3181 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3241 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3301 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3361 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3421 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3481 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     3541 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3601 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3661 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3721 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3781 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3841 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3901 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3961 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4021 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4081 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4141 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4201 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4261 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4321 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4381 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4441 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4501 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4561 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4621 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4681 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4741 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4801 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4861 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4921 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4981 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5041 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5101 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5161 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5221 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5281 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5341 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5401 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5461 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5521 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5581 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5641 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5701 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5761 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5821 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5881 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     5941 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     6001 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
     6061 tacgaattaa ttcgagctcg ccc
//
