LOCUS       sequence-448252-        7086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The ORF of human SMAD4, variant 1 cDNA amplified from the RT human 
            liver RNA was cloned at the NheI/HindIII sites of the pcDNA3.1(+), 
            the mammalian expression vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7086)
  AUTHORS   Zolfaghari R, Ross C
  TITLE     Zolfaghari and Ross plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7086)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3.1_hSMAD4
FEATURES             Location/Qualifiers
     source          1..7086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2677..2694)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2683..2907
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2953..3381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3040..3059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3250..3271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3390..3410)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3395..3724
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3575..3710
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3637..3656
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3791..4585
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3845..3864)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4455..4474
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4759..4880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4796..4815)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4850..4869
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4929..4945)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4929..4945)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4942..4964)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4953..4969
                     /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(4977..4983,4984..5001,5002..5007))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5022..5043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5160..5177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5331..5919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5411..5430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6090..6881,6882..6950))
                     /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     6713..6732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6951..7055)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatgg acaatatgtc tattacgaat acaccaacaa gtaatgatgc ctgtctgagc
      961 attgtgcata gtttgatgtg ccatagacaa ggtggagaga gtgaaacatt tgcaaaaaga
     1021 gcaattgaaa gtttggtaaa gaagctgaag gagaaaaaag atgaattgga ttctttaata
     1081 acagctataa ctacaaatgg agctcatcct agtaaatgtg ttaccataca gagaacattg
     1141 gatgggaggc ttcaggtggc tggtcggaaa ggatttcctc atgtgatcta tgcccgtctc
     1201 tggaggtggc ctgatcttca caaaaatgaa ctaaaacatg ttaaatattg tcagtatgcg
     1261 tttgacttaa aatgtgatag tgtctgtgtg aatccatatc actacgaacg agttgtatca
     1321 cctggaattg atctctcagg attaacactg cagagtaatg ctccatcaag tatgatggtg
     1381 aaggatgaat atgtgcatga ctttgaggga cagccatcgt tgtccactga aggacattca
     1441 attcaaacca tccagcatcc accaagtaat cgtgcatcga cagagacata cagcacccca
     1501 gctctgttag ccccatctga gtctaatgct accagcactg ccaactttcc caacattcct
     1561 gtggcttcca caagtcagcc tgccagtata ctggggggca gccatagtga aggactgttg
     1621 cagatagcat cagggcctca gccaggacag cagcagaatg gatttactgg tcagccagct
     1681 acttaccatc ataacagcac taccacctgg actggaagta ggactgcacc atacacacct
     1741 aatttgcctc accaccaaaa cggccatctt cagcaccacc cgcctatgcc gccccatccc
     1801 ggacattact ggcctgttca caatgagctt gcattccagc ctcccatttc caatcatcct
     1861 gctcctgagt attggtgttc cattgcttac tttgaaatgg atgttcaggt aggagagaca
     1921 tttaaggttc cttcaagctg ccctattgtt actgttgatg gatacgtgga cccttctgga
     1981 ggagatcgct tttgtttggg tcaactctcc aatgtccaca ggacagaagc cattgagaga
     2041 gcaaggttgc acataggcaa aggtgtgcag ttggaatgta aaggtgaagg tgatgtttgg
     2101 gtcaggtgcc ttagtgacca cgcggtcttt gtacagagtt actacttaga cagagaagct
     2161 gggcgtgcac ctggagatgc tgttcataag atctacccaa gtgcatatat aaaggtcttt
     2221 gatttgcgtc agtgtcatcg acagatgcag cagcaggcgg ctactgcaca agctgcagca
     2281 gctgcccagg cagcagccgt ggcaggaaac atccctggcc caggatcagt aggtggaata
     2341 gctccagcta tcagtctgtc agctgctgct ggaattggtg ttgatgacct tcgtcgctta
     2401 tgcatactca ggatgagttt tgtgaaaggc tggggaccgg attacccaag acagagcatc
     2461 aaagaaacac cttgctggat tgaaattcac ttacaccggg ccctccagct cctagacgaa
     2521 gtacttcata ccatgccgat tgcagaccca caacctttag actgaaagct tggtaccgag
     2581 ctcggatcca ctagtccagt gtggtggaat tctgcagata tccagcacag tggcggccgc
     2641 tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     2701 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2761 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2821 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2881 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2941 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3001 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3061 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3121 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3181 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3241 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3301 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3361 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3421 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3481 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3541 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3601 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3661 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3721 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     3781 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3841 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3901 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3961 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4021 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4081 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4141 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4201 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4261 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4321 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4381 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4441 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4501 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4561 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4621 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4681 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4741 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4801 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4861 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4921 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4981 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5041 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5101 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5161 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5221 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5281 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5341 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5401 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5461 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5521 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5581 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5641 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5701 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5761 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5821 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5881 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5941 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6001 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6061 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6121 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6181 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6241 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6301 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6361 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6421 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6481 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6541 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6601 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6661 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6721 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6781 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6841 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6901 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6961 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7021 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7081 gacgtc
//
