LOCUS       sequence-152151-        6250 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 6250)
  AUTHORS   Liu F, Hata A, Baker JC, Doody J, Carcamo J, Harland RM, Massague J
  TITLE     A human Mad protein acting as a BMP-regulated transcriptional 
            activator.
  JOURNAL   Nature. 1996 Jun 13. 381(6583):620-3.
  PUBMED    8637600
REFERENCE   2  (bases 1 to 6250)
  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. 
            1996 Jun 13. 381(6583):620-3."
COMMENT     Sequence Label: pCMV5 Flag-Smad1
FEATURES             Location/Qualifiers
     source          1..6250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..62
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     54..71
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     55..71
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        232..611
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        612..815
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     766..786
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             875..898
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2505..3127
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2704..2723)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3151..3171)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3156..3485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3336..3471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3523..3542)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3524..3542)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3556..3572)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3556..3572)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3569..3591)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3580..3596
                     /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(3604..3610,3611..3628,3629..3634))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3649..3670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3787..3804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3958..4546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4038..4057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4717..5508,5509..5577))
                     /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     5340..5359
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5578..5682)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5709..6164
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5796..5815)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6006..6027
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
       61 cgacggccag tgccaagctg atctatacat tgaatcaata ttggcaatta gccatattag
      121 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
      181 atcataatat gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat
      241 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      301 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      361 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      421 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      481 atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      541 cccagtacat gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg
      601 ctattaccat ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact
      661 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      721 atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta
      781 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagaattcag
      841 atctggtacc acgcgtatcg ataagcttac catggactac aaggacgacg atgacaagaa
      901 tgtgacaagt ttattttcct ttacaagtcc agctgtgaag agacttcttg ggtggaaaca
      961 gggcgatgaa gaagaaaaat gggcagagaa agctgttgat gctttggtga aaaaactgaa
     1021 gaaaaagaaa ggtgccatgg aggaactgga aaaggccttg agctgcccag ggcaaccgag
     1081 taactgtgtc accattcccc gctctctgga tggcaggctg caagtctccc accggaaggg
     1141 actgcctcat gtcatttact gccgtgtgtg gcgctggccc gatcttcaga gccaccatga
     1201 actaaaacca ctggaatgct gtgagtttcc ttttggttcc aagcagaagg aggtctgcat
     1261 caatccctac cactataaga gagtagaaag ccctgtactt cctcctgtgc tggttccaag
     1321 acacagcgaa tataatcctc agcacagcct cttagctcag ttccgtaact taggacaaaa
     1381 tgagcctcac atgccactca acgccacttt tccagattct ttccagcaac ccaacagcca
     1441 cccgtttcct cactctccca atagcagtta cccaaactct cctgggagca gcagcagcac
     1501 ctaccctcac tctcccacca gctcagaccc aggaagccct ttccagatgc cagctgatac
     1561 gcccccacct gcttacctgc ctcctgaaga ccccatgacc caggatggct ctcagccgat
     1621 ggacacaaac atgatggcgc ctcccctgcc ctcagaaatc aacagaggag atgttcaggc
     1681 ggttgcttat gaggaaccaa aacactggtg ctctattgtc tactatgagc tcaacaatcg
     1741 tgtgggtgaa gcgttccatg cctcctccac aagtgtgttg gtggatggtt tcactgatcc
     1801 ttccaacaat aagaaccgtt tctgccttgg gctgctctcc aatgttaacc ggaattccac
     1861 tattgaaaac accaggcggc atattggaaa aggagttcat ctttattatg ttggagggga
     1921 ggtgtatgcc gaatgcctta gtgacagtag catctttgtg caaagtcgga actgcaacta
     1981 ccatcatgga tttcatccta ctactgtttg caagatccct agtgggtgta gtctgaaaat
     2041 ttttaacaac caagaatttg ctcagttatt ggcacagtct gtgaaccatg gatttgagac
     2101 agtctatgag cttacaaaaa tgtgtactat acgtatgagc tttgtgaagg gctggggagc
     2161 agaataccac cgccaggatg ttactagcac cccctgctgg attgagatac atctgcacgg
     2221 ccccctccag tggctggata aagttcttac tcaaatgggt tcacctcata atcctatttc
     2281 atctgtatct taaatggccc caggcatctg cctctggaaa actattgagc cttgcatgta
     2341 cttgaaggat ggatgagtca gacacgattg agaactgaca aaggagcctt gataatactt
     2401 gacctctgtg accaactgtt ggattcagaa atttaaacaa aaaaaaaaaa aaaaaaaggg
     2461 cggccgcaag cttgcatgcc tgcaggtcga ctctagagga tcccgggtgg catccctgtg
     2521 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     2581 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     2641 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     2701 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     2761 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     2821 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     2881 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     2941 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta aaataactat
     3001 accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa ccggtgggac
     3061 atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag tagatgcctg
     3121 ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3301 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3361 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3421 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3481 gcaaaaagct cctcgaggaa ctgaaaaacc agaaagttaa ttccctatag tgagtcgtat
     3541 taaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3601 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3661 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3721 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3781 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3841 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3901 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3961 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4021 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4081 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4141 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4201 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4261 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4321 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4381 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4441 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4501 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4561 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4621 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4681 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4741 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4801 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4861 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4921 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4981 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5041 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5101 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5161 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5221 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5281 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5341 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5401 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5461 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5521 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5581 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5641 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5701 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5761 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5821 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5881 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5941 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6001 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     6061 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     6121 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg
     6181 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     6241 aaagggggat
//
