LOCUS       sequence-217111-        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"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1743..1766
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2115..2144
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2163..2186
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2206..2222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2206..2222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2233..2855
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2432..2451)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(3004..3592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3084..3103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3921..3968
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3993..4012
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4200..4994)
                     /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(4200..4994)
                     /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(4311..4330)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4921..4940
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5029..5386)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5043..5178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5195..5386
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5343..5363
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5388..5492)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5519..5947
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5606..5625)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5816..5837
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcatcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat gcacgcccac tgcctgccct tccttctgca cgcctggtgg gccctactcc
     1081 aggcgggtgc tgcgacggtg gccactgcgc tcctgcgtac gcgggggcag ccctcgtcgc
     1141 catcccctct ggcgtacatg ctgagcctct accgcgaccc gctgccgagg gcagacatca
     1201 tccgcagcct acaggcagaa gatgtggcag tggatgggca gaactggacg tttgcttttg
     1261 acttctcctt cctgagccaa caagaggatc tggcatgggc tgagctccgg ctgcagctgt
     1321 ccagccctgt ggacctcccc actgagggct cacttgccat tgagattttc caccagccaa
     1381 agcccgacac agagcaggct tcagacagct gcttagagcg gtttcagatg gacctattca
     1441 ctgtcacttt gtcccaggtc accttttcct tgggcagcat ggttttggag gtgaccaggc
     1501 ctctctccaa gtggctgaag caccctgggg ccctggagaa gcagatgtcc agggtagctg
     1561 gagagtgctg gccgcggccc cccacaccgc ctgccaccaa tgtgctcctt atgctctact
     1621 ccaacctctc gcaggancag angcagctgg gtgggtccac cttgctgtgg gaagccgaga
     1681 gctcctggcg ggcccaggag ggacagctgt cctgggagtg gggcaagagg caccgtcgac
     1741 atgattacaa ggatgacgac gataagcatc acttgccaga cagaagtcaa ctgtgtcgga
     1801 aggtcaagtt ccaggtggac ttcaacctga tcggatgggg ctcctggatc atctacccca
     1861 agcagtacaa cgcctatcgc tgtgagggcg agtgtcctaa tcctgttggg gaggagtttc
     1921 atccgaccaa ccatgcatac atccagagtc tgctgaaacg ttaccagccc caccgagtcc
     1981 cttccacttc ttgtgcccca gtgaagacca agccgctgag catgctgtat gtggataatg
     2041 gcagagtgct cctagatcac cataaagaca tgatcgtgga agaatgtggg tgcctctgac
     2101 gtacgcggcc gctcgagcag aaactcatct cagaagagga tctggcagca aatgatatcc
     2161 tggattacaa ggatgacgac gataaggttt aaacggccgg ccgcggtcat agctgtttcc
     2221 tgaacagatc ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     2281 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     2341 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     2401 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     2461 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     2521 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     2581 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     2641 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     2701 cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg
     2761 ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca
     2821 tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc ggcgagcggt
     2881 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2941 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3001 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3061 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3121 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3181 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3241 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3301 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3361 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3421 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3481 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3541 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3601 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3661 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3721 taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac
     3781 gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga
     3841 aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg
     3901 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt
     3961 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     4021 ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     4081 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     4141 atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     4201 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     4261 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     4321 gtagccaacg ctatgtcctg atagcgatcc gccacaccca gccggccaca gtcgatgaat
     4381 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     4441 acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg
     4501 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     4561 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     4621 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     4681 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     4741 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     4801 gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     4861 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     4921 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     4981 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc
     5041 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     5101 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     5161 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     5221 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     5281 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     5341 acaccctaac tgacacacat tccacagctg gttctttccg cctcaggact cttccttttt
     5401 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5461 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5521 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5581 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5641 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5701 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5761 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5821 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5881 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5941 gcgaatttt
//
