LOCUS       sequence-372918-        6058 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 6058)
  AUTHORS   Eppert K, Scherer SW, Ozcelik H, Pirone R, Hoodless P, Kim H, Tsui 
            LC, Bapat B, Gallinger S, Andrulis IL, Thomsen GH, Wrana JL, 
            Attisano L
  TITLE     MADR2 maps to 18q21 and encodes a TGFbeta-regulated MAD-related 
            protein that is functionally mutated in colorectal carcinoma.
  JOURNAL   Cell. 1996 Aug 23. 86(4):543-52.
  PUBMED    8752209
REFERENCE   2  (bases 1 to 6058)
  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: "Cell. 1996 
            Aug 23. 86(4):543-52."
COMMENT     Sequence Label: pCMV5B-HA-Smad2
FEATURES             Location/Qualifiers
     source          1..6058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(78..98)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        83..412
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      263..398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(450..469)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(451..469)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(483..499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(483..499)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(496..518)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    507..523
                     /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(531..537,538..555,556..561))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    576..597
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(885..1473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(965..984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1644..2435,2436..2504))
                     /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     2267..2286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2505..2609)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2636..3091
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2723..2742)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2933..2954
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3217..3239
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3231..3248
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3232..3248
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3409..3788
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3789..3992
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3943..3963
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4044..4070
                     /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    join(5490..6058,1..54)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5689..5708)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac
       61 gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
      121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
      181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
      241 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
      301 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
      361 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcct
      421 cgaggaactg aaaaaccaga aagttaattc cctatagtga gtcgtattaa attcgtaatc
      481 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
      541 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
      601 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
      661 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
      721 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
      781 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
      841 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      901 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      961 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1021 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1081 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1141 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1201 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1261 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1321 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1381 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     1441 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     1501 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     1561 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1621 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1681 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1741 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     1801 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     1861 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     1921 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     1981 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2041 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2101 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2161 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2221 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2281 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2341 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     2401 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     2461 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     2521 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     2581 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     2641 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2701 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2761 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2821 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2881 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2941 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3001 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3061 aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt
     3121 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     3181 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     3241 cggccagtgc caagctgatc tatacattga atcaatattg gcaattagcc atattagtca
     3301 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     3361 ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttga cattgattat
     3421 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     3481 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3541 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3601 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3661 tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     3721 agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     3781 ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac
     3841 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     3901 aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc
     3961 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag aattgatctg
     4021 gtaccacgcg tatcgatacc atgtatgatg ttcctgatta tgctagcctc ggtcgacaat
     4081 cgtccatctt gccattcacg ccgccagttg tgaagagact gctgggatgg aagaagtcag
     4141 ctggtgggtc tggaggagca ggcggaggag agcagaatgg gcaggaagaa aagtggtgtg
     4201 agaaagcagt gaaaagtctg gtgaagaagc taaagaaaac aggacgatta gatgagcttg
     4261 agaaagccat caccactcaa aactgtaata ctaaatgtgt taccatacca agcacttgct
     4321 ctgaaatttg gggactgagt acaccaaata cgatagatca gtgggataca acaggccttt
     4381 acagcttctc tgaacaaacc aggtctcttg atggtcgtct ccaggtatcc catcgaaaag
     4441 gattgccaca tgttatatat tgccgattat ggcgctggcc tgatcttcac agtcatcatg
     4501 aactcaaggc aattgaaaac tgcgaatatg cttttaatct taaaaaggat gaagtatgtg
     4561 taaaccctta ccactatcag agagttgaga caccagtttt gcctccagta ttagtgcccc
     4621 gacacaccga gatcctaaca gaacttccgc ctctggatga ctatactcac tccattccag
     4681 aaaacactaa cttcccagca ggaattgagc cacagagtaa ttatattcca gaaacgccac
     4741 ctcctggata tatcagtgaa gatggagaaa caagtgacca acagttgaat caaagtatgg
     4801 acacaggctc tccagcagaa ctatctccta ctactctttc ccctgttaat catagcttgg
     4861 atttacagcc agttacttac tcagaacctg cattttggtg ttcgatagca tattatgaat
     4921 taaatcagag ggttggagaa accttccatg catcacagcc ctcactcact gtagatggct
     4981 ttacagaccc atcaaattca gagaggttct gcttaggttt actctccaat gttaaccgaa
     5041 atgccacggt agaaatgaca agaaggcata taggaagagg agtgcgctta tactacatag
     5101 gtggggaagt ttttgctgag tgcctaagtg atagtgcaat ctttgtgcag agccccaatt
     5161 gtaatcagag atatggctgg caccctgcaa cagtgtgtaa aattccacca ggctgtaatc
     5221 tgaagatctt caacaaccag gaatttgctg ctcttctggc tcagtctgtt aatcagggtt
     5281 ttgaagccgt ctatcagcta actagaatgt gcaccataag aatgagtttt gtgaaagggt
     5341 ggggagcaga ataccgaagg cagacggtaa caagtactcc ttgctggatt gaacttcatc
     5401 tgaatggacc tctacagtgg ttggacaaag tattaactca gatgggatcc ccttcagtgc
     5461 gttgctcaag catgtcataa tctagacccg ggtggcatcc ctgtgacccc tccccagtgc
     5521 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     5581 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     5641 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     5701 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     5761 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     5821 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     5881 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     5941 gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag caggaggacg
     6001 tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg agttgctt
//
