LOCUS       Exported                6058 bp ds-DNA     circular SYN 23-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCMV5B-HA-Smad2
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 Sep 23, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6058
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     polyA_signal    2082..2704
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2281..2300)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2728..2748)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2733..3062
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2913..3048
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2975..2994
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3100..3119)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3101..3119)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3133..3149)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3133..3149)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3146..3168)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3157..3173
                     /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(3181..3211)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3226..3247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3364..3381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3535..4123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3615..3634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4294..5154)
                     /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     4917..4936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5155..5259)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5286..5741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5373..5392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5583..5604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5867..5889
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5881..5898
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5882..5898
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgatc tggtaccacg cgtatcgata ccatgtatga tgttcctgat tatgctagcc
      661 tcggtcgaca atcgtccatc ttgccattca cgccgccagt tgtgaagaga ctgctgggat
      721 ggaagaagtc agctggtggg tctggaggag caggcggagg agagcagaat gggcaggaag
      781 aaaagtggtg tgagaaagca gtgaaaagtc tggtgaagaa gctaaagaaa acaggacgat
      841 tagatgagct tgagaaagcc atcaccactc aaaactgtaa tactaaatgt gttaccatac
      901 caagcacttg ctctgaaatt tggggactga gtacaccaaa tacgatagat cagtgggata
      961 caacaggcct ttacagcttc tctgaacaaa ccaggtctct tgatggtcgt ctccaggtat
     1021 cccatcgaaa aggattgcca catgttatat attgccgatt atggcgctgg cctgatcttc
     1081 acagtcatca tgaactcaag gcaattgaaa actgcgaata tgcttttaat cttaaaaagg
     1141 atgaagtatg tgtaaaccct taccactatc agagagttga gacaccagtt ttgcctccag
     1201 tattagtgcc ccgacacacc gagatcctaa cagaacttcc gcctctggat gactatactc
     1261 actccattcc agaaaacact aacttcccag caggaattga gccacagagt aattatattc
     1321 cagaaacgcc acctcctgga tatatcagtg aagatggaga aacaagtgac caacagttga
     1381 atcaaagtat ggacacaggc tctccagcag aactatctcc tactactctt tcccctgtta
     1441 atcatagctt ggatttacag ccagttactt actcagaacc tgcattttgg tgttcgatag
     1501 catattatga attaaatcag agggttggag aaaccttcca tgcatcacag ccctcactca
     1561 ctgtagatgg ctttacagac ccatcaaatt cagagaggtt ctgcttaggt ttactctcca
     1621 atgttaaccg aaatgccacg gtagaaatga caagaaggca tataggaaga ggagtgcgct
     1681 tatactacat aggtggggaa gtttttgctg agtgcctaag tgatagtgca atctttgtgc
     1741 agagccccaa ttgtaatcag agatatggct ggcaccctgc aacagtgtgt aaaattccac
     1801 caggctgtaa tctgaagatc ttcaacaacc aggaatttgc tgctcttctg gctcagtctg
     1861 ttaatcaggg ttttgaagcc gtctatcagc taactagaat gtgcaccata agaatgagtt
     1921 ttgtgaaagg gtggggagca gaataccgaa ggcagacggt aacaagtact ccttgctgga
     1981 ttgaacttca tctgaatgga cctctacagt ggttggacaa agtattaact cagatgggat
     2041 ccccttcagt gcgttgctca agcatgtcat aatctagacc cgggtggcat ccctgtgacc
     2101 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     2161 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     2221 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     2281 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     2341 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     2401 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     2461 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     2521 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc
     2581 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     2641 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     2701 aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2761 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2821 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2881 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2941 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3001 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3061 aaaagctcct cgaggaactg aaaaaccaga aagttaattc cctatagtga gtcgtattaa
     3121 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3181 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3241 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3301 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3961 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4081 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4141 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4201 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4261 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4321 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4381 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4441 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4501 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4561 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4621 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4681 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4741 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4801 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4861 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4921 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4981 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5041 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5101 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5161 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5221 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5281 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5341 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5401 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5461 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5521 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5581 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5641 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5701 atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg
     5761 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     5821 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     5881 tgtaaaacga cggccagtgc caagctgatc tatacattga atcaatattg gcaattagcc
     5941 atattagtca ttggttatat agcataaatc aatattggct attggccatt gcatacgttg
     6001 tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc gccatgtt
//
