LOCUS       sequence-152156-        6082 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 6082)
  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 6082)
  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 Smad1-HA
FEATURES             Location/Qualifiers
     source          1..6082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     82..104
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     96..113
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     97..113
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        274..653
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        654..857
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     808..828
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2336..2362
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2342..2368
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     polyA_signal    2379..3001
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2578..2597)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3025..3045)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3030..3359
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3210..3345
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3397..3416)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3398..3416)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3430..3446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3430..3446)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3443..3465)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3454..3470
                     /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(3478..3484,3485..3502,3503..3508))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3523..3544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3661..3678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3832..4420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3912..3931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4591..5382,5383..5451))
                     /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     5214..5233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5452..5556)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5583..6038
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5670..5689)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5880..5901
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
       61 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
      121 tgatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
      181 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
      241 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat
      301 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      361 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      421 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      481 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
      541 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
      601 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      661 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      721 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      781 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
      841 tctatataag cagagctcgt ttagtgaacc gtcagaattc agatctggta ccacgcgtat
      901 cgataagctt tatccaagga gtataactag tgctgtcatt atgaatgtga caagtttatt
      961 ttcctttaca agtccagctg tgaagagact tcttgggtgg aaacagggcg atgaagaaga
     1021 aaaatgggca gagaaagctg ttgatgcttt ggtgaaaaaa ctgaagaaaa agaaaggtgc
     1081 catggaggaa ctggaaaagg ccttgagctg cccagggcaa ccgagtaact gtgtcaccat
     1141 tccccgctct ctggatggca ggctgcaagt ctcccaccgg aagggactgc ctcatgtcat
     1201 ttactgccgt gtgtggcgct ggcccgatct tcagagccac catgaactaa aaccactgga
     1261 atgctgtgag tttccttttg gttccaagca gaaggaggtc tgcatcaatc cctaccacta
     1321 taagagagta gaaagccctg tacttcctcc tgtgctggtt ccaagacaca gcgaatataa
     1381 tcctcagcac agcctcttag ctcagttccg taacttagga caaaatgagc ctcacatgcc
     1441 actcaacgcc acttttccag attctttcca gcaacccaac agccacccgt ttcctcactc
     1501 tcccaatagc agttacccaa actctcctgg gagcagcagc agcacctacc ctcactctcc
     1561 caccagctca gacccaggaa gccctttcca gatgccagct gatacgcccc cacctgctta
     1621 cctgcctcct gaagacccca tgacccagga tggctctcag ccgatggaca caaacatgat
     1681 ggcgcctccc ctgccctcag aaatcaacag aggagatgtt caggcggttg cttatgagga
     1741 accaaaacac tggtgctcta ttgtctacta tgagctcaac aatcgtgtgg gtgaagcgtt
     1801 ccatgcctcc tccacaagtg tgttggtgga tggtttcact gatccttcca acaataagaa
     1861 ccgtttctgc cttgggctgc tctccaatgt taaccggaat tccactattg aaaacaccag
     1921 gcggcatatt ggaaaaggag ttcatcttta ttatgttgga ggggaggtgt atgccgaatg
     1981 ccttagtgac agtagcatct ttgtgcaaag tcggaactgc aactaccatc atggatttca
     2041 tcctactact gtttgcaaga tccctagtgg gtgtagtctg aaaattttta acaaccaaga
     2101 atttgctcag ttattggcac agtctgtgaa ccatggattt gagacagtct atgagcttac
     2161 aaaaatgtgt actatacgta tgagctttgt gaagggctgg ggagcagaat accaccgcca
     2221 ggatgttact agcaccccct gctggattga gatacatctg cacggccccc tccagtggct
     2281 ggataaagtt cttactcaaa tgggttcacc tcataatcct atttcatctg tatcttatcc
     2341 gtatgatgtt cctgattatg ctagtctcta gggatcccgg gtggcatccc tgtgacccct
     2401 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     2461 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     2521 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     2581 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     2641 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     2701 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     2761 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     2821 tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc
     2881 aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga
     2941 gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat
     3001 tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3121 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3181 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3241 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3301 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3361 agctcctcga ggaactgaaa aaccagaaag ttaattccct atagtgagtc gtattaaatt
     3421 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3481 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3541 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3601 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3661 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3721 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3781 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3841 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3901 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3961 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4021 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4081 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4141 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4201 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4261 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4321 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4381 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4441 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4501 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4561 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4621 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4681 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4741 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4801 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4861 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4921 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4981 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5041 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5101 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5161 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5221 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5281 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5341 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5401 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5461 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5521 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5581 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5641 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5701 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5761 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5821 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5881 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5941 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     6001 taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc
     6061 aactgttggg aagggcgatc gg
//
