LOCUS       sequence-222465-        5949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of the human NODAL open reading frame (with
            a mutated Cysteine residue) with an internal DYK tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5949)
  AUTHORS   Findlay SD, Bilyk O, Lypka K, Waskiewicz AJ, Postovit L
  TITLE     Genetically regulated human NODAL splice variants are differentially
            post-transcriptionally processed and functionally distinct
  JOURNAL   (2018) bioRxiv 276170
REFERENCE   2  (bases 1 to 5949)
  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: "(2018) 
            bioRxiv 276170"
COMMENT     Sequence Label: NODAL-matDYK-C312S
FEATURES             Location/Qualifiers
     source          1..5949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        162..541
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        542..745
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     696..716
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     771..790
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        771..789
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1562..1585
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1934..1963
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1982..2005
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2025..2041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2025..2041)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2052..2674
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2251..2270)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2823..3411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2903..2922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3740..3787
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3812..3831
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4019..4813)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4019..4813)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4130..4149)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4740..4759
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4848..5205)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4862..4997
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5014..5205
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5162..5182
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5207..5311)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5338..5793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5425..5444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5635..5656
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5919..5941
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5933..5949,1)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(5934..5949,1)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgccaagctg atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta
       61 tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat
      121 gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat tattgactag
      181 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      241 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      301 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      361 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      421 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      481 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      541 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
      601 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      661 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
      721 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagaattttg taatacgact
      781 cactataggg cggccgggaa ttcatcgact ggatccggta ccgaggagat ctgccgccgc
      841 gatcgccatg cacgcccact gcctgccctt ccttctgcac gcctggtggg ccctactcca
      901 ggcgggtgct gcgacggtgg ccactgcgct cctgcgtacg cgggggcagc cctcgtcgcc
      961 atcccctctg gcgtacatgc tgagcctcta ccgcgacccg ctgccgaggg cagacatcat
     1021 ccgcagccta caggcagaag atgtggcagt ggatgggcag aactggacgt ttgcttttga
     1081 cttctccttc ctgagccaac aagaggatct ggcatgggct gagctccggc tgcagctgtc
     1141 cagccctgtg gacctcccca ctgagggctc acttgccatt gagattttcc accagccaaa
     1201 gcccgacaca gagcaggctt cagacagctg cttagagcgg tttcagatgg acctattcac
     1261 tgtcactttg tcccaggtca ccttttcctt gggcagcatg gttttggagg tgaccaggcc
     1321 tctctccaag tggctgaagc accctggggc cctggagaag cagatgtcca gggtagctgg
     1381 agagtgctgg ccgcggcccc ccacaccgcc tgccaccaat gtgctcctta tgctctactc
     1441 caacctctcg caggagcaga ggcagctggg tgggtccacc ttgctgtggg aagccgagag
     1501 ctcctggcgg gcccaggagg gacagctgtc ctgggagtgg ggcaagaggc accgtcgaca
     1561 tgattacaag gatgacgacg ataagcatca cttgccagac agaagtcaac tgtgtcggaa
     1621 ggtcaagttc caggtggact tcaacctgat cggatggggc tcctggatca tctaccccaa
     1681 gcagtacaac gcctatcgct gtgagggcga gtgtcctaat cctgttgggg aggagtttca
     1741 tccgaccaac catgcataca tccagagtct gctgaaacgt taccagcccc accgagtccc
     1801 ttccacttct tgtgccccag tgaagaccaa gccgctgagc atgctgtatg tggataatgg
     1861 cagagtgctc ctagatcacc ataaagacat gatcgtggaa gaatgtgggt gcctctgacg
     1921 tacgcggccg ctcgagcaga aactcatctc agaagaggat ctggcagcaa atgatatcct
     1981 ggattacaag gatgacgacg ataaggttta aacggccggc cgcggtcata gctgtttcct
     2041 gaacagatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     2101 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     2161 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     2221 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     2281 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     2341 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     2401 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     2461 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     2521 ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc
     2581 tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat
     2641 gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagcg gcgagcggta
     2701 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2761 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2821 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2881 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2941 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3001 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3061 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3121 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3181 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3241 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3301 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3361 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3421 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3481 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3541 aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc cgccccgacg
     3601 ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa
     3661 acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     3721 gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt
     3781 attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc
     3841 cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     3901 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     3961 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
     4021 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     4081 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     4141 tagccaacgc tatgtcctga tagcgatccg ccacacccag ccggccacag tcgatgaatc
     4201 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     4261 cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga
     4321 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     4381 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     4441 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     4501 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     4561 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     4621 ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     4681 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     4741 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     4801 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct
     4861 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg
     4921 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
     4981 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga
     5041 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct
     5101 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     5161 caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc ttcctttttc
     5221 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5281 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5341 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5401 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5461 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5521 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5581 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5641 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5701 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5761 cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg
     5821 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     5881 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     5941 gacggccag
//
