LOCUS       sequence-197586-        5929 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 5929)
  AUTHORS   Labbe E, Silvestri C, Hoodless PA, Wrana JL, Attisano L
  TITLE     Smad2 and Smad3 positively and negatively regulate TGF 
            beta-dependent transcription through the forkhead DNA-binding 
            protein FAST2.
  JOURNAL   Mol Cell. 1998 Jul . 2(1):109-20.
  PUBMED    9702197
REFERENCE   2  (bases 1 to 5929)
  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: "Mol Cell. 
            1998 Jul . 2(1):109-20."
COMMENT     Sequence Label: pCMV5B-Flag-Smad3
FEATURES             Location/Qualifiers
     source          1..5929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             953..976
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2270..2892
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2469..2488)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2916..2936)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2921..3250
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3101..3236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3288..3307)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3289..3307)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3321..3337)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3321..3337)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3334..3356)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3345..3361
                     /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)."
     promoter        complement(join(3369..3375,3376..3393,3394..3399))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3414..3435
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3552..3569)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3723..4311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3803..3822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4482..5273,5274..5342))
                     /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     5105..5124
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5343..5447)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5474..5902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5561..5580)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5771..5792
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctgg taccacgcgt atcgatacca tggactacaa
      961 ggacgacgat gacaagggtc gactgtcgtc catcctgcct ttcactcccc cgatcgtgaa
     1021 gcgcctgctg ggctggaaga agggcgagca gaacgggcag gaggagaaat ggtgcgagaa
     1081 ggcggtcaag agcctggtca agaaactcaa gaagacgggg cagctggacg agctggagaa
     1141 ggccatcacc acgcagaacg tcaacaccaa gtgcatcacc atccccaggt ccctggatgg
     1201 ccggttgcag gtgtcccatc ggaaggggct ccctcatgtc atctactgcc gcctgtggcg
     1261 atggccagac ctgcacagcc accacgagct gcgggccatg gagctgtgtg agttcgcctt
     1321 caatatgaag aaggacgagg tctgcgtgaa tccctaccac taccagagag tagagacacc
     1381 agttctacct cctgtgttgg tgccacgcca cacagagatc ccggccgagt tccccccact
     1441 ggacgactac agccattcca tccccgaaaa cactaacttc cccgcaggca tcgagcccca
     1501 gagcaatatt ccagagaccc caccccctgg ctacctgagt gaagatggag aaaccagtga
     1561 ccaccagatg aaccacagca tggacgcagg ttctccaaac ctatccccga atccgatgtc
     1621 cccagcacat aataacttgg acctgcagcc agttacctac tgcgagccgg ccttctggtg
     1681 ctccatctcc tactacgagc tgaaccagcg cgtcggggag acattccacg cctcgcagcc
     1741 atccatgact gtggatggct tcaccgaccc ctccaattcg gagcgcttct gcctagggct
     1801 gctctccaat gtcaacagga atgcagcagt ggagctgaca cggagacaca tcggaagagg
     1861 cgtgcggctc tactacatcg gaggggaggt cttcgcagag tgcctcagtg acagcgctat
     1921 ttttgtccag tctcccaact gtaaccagcg ctatggctgg cacccggcca ccgtctgcaa
     1981 gatcccacca ggatgcaacc tgaagatctt caacaaccag gagttcgctg ccctcctggc
     2041 ccagtcggtc aaccagggct ttgaggctgt ctaccagttg acccgaatgt gcaccatccg
     2101 catgagcttc gtcaaaggct ggggagcgga gtacaggaga cagactgtga ccagtacccc
     2161 ctgctggatt gagctgcacc tgaatgggcc tttgcagtgg cttgacaagg tcctcaccca
     2221 gatgggctcc ccaagcatcc gctgttccag tgtgtcttag tctagacccg ggtggcatcc
     2281 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     2341 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     2401 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     2461 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     2521 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     2581 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     2641 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     2701 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata
     2761 actataccag caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt
     2821 gggacatttg agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat
     2881 gcctgttgaa ttgggtacgc ggccagcttg gctgtggaat gtgtgtcagt tagggtgtgg
     2941 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3001 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3061 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3121 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3181 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3241 cttttgcaaa aagctcctcg aggaactgaa aaaccagaaa gttaattccc tatagtgagt
     3301 cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3361 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3421 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3481 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3541 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3601 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3661 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3721 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3781 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3841 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3901 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3961 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4021 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4081 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4141 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4201 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4261 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4321 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4381 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4441 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4501 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4561 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4621 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4681 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4741 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4801 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4861 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4921 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4981 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5041 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5101 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5161 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5221 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5281 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5341 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5401 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5461 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5521 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5581 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5641 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5701 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5761 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     5821 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     5881 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattt
//
