LOCUS       sequence-222242-        5895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of a human NODAL splice variant open 
            reading frame (with a mutated Cysteine residue) with a C-terminal 
            MYC-DYK tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5895)
  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 5895)
  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: NODALvar-MYC-DYK-C302S
FEATURES             Location/Qualifiers
     source          1..5895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        533..736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     687..707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     762..781
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        762..780
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1871..1900
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1919..1942
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1962..1978)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1962..1978)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1989..2611
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2188..2207)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2760..3348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2840..2859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3677..3724
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3749..3768
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3956..4750)
                     /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(3956..4750)
                     /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(4067..4086)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4677..4696
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4785..5142)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4799..4934
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4951..5142
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5099..5119
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5144..5248)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5275..5730
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5362..5381)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5572..5593
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5856..5878
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5870..5887
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5871..5887
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata
       61 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta
      121 tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata
      181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      361 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
      421 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
      481 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      541 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      601 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      661 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
      721 ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac tcactatagg
      781 gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg cgatcgccat
      841 gcacgcccac tgcctgccct tccttctgca cgcctggtgg gccctactcc aggcgggtgc
      901 tgcgacggtg gccactgcgc tcctgcgtac gcgggggcag ccctcgtcgc catcccctct
      961 ggcgtacatg ctgagcctct accgcgaccc gctgccgagg gcagacatca tccgcagcct
     1021 acaggcagaa gatgtggcag tggatgggca gaactggacg tttgcttttg acttctcctt
     1081 cctgagccaa caagaggatc tggcatgggc tgagctccgg ctgcagctgt ccagccctgt
     1141 ggacctcccc actgagggct cacttgccat tgagattttc caccagccaa agcccgacac
     1201 agagcaggct tcagacagct gcttagagcg gtttcagatg gacctattca ctgtcacttt
     1261 gtcccaggtc accttttcct tgggcagcat ggttttggag gtgaccaggc ctctctccaa
     1321 gtggctgaag caccctgggg ccctggagaa gcagatgtcc agggtagctg gagagtgctg
     1381 gccgcggccc cccacaccgc ctgccaccaa tgtgctcctt atgctctact ccaacctctc
     1441 gcaggagcag aggcagctgg gtgggtccac cttgctgtgg gaagccgaga gctcctggcg
     1501 ggcccaggag ggacagctgt cctgggagtg gggcaagagg caccgtcgac atcacttgcc
     1561 agacagaagt caactgtgtc ggaaggtcaa gttccaggtg gacttcaacc tgatcggatg
     1621 gggctcctgg atcatctacc ccaagcagta caacgcctat cgctgtgagg gcgagtgtcc
     1681 taatcctgtt ggggaggagt ttcatccgac caaccatgca tacatccagg tggccctgcc
     1741 ctcctgtcca aggtcatatg ggaccaaaat gttttcattt tactccatga agtctggaat
     1801 gagaatttct tggacttgca atatatcctc catgccaagc ctcagagtct gcacgcgtac
     1861 gcggccgctc gagcagaaac tcatctcaga agaggatctg gcagcaaatg atatcctgga
     1921 ttacaaggat gacgacgata aggtttaaac ggccggccgc ggtcatagct gtttcctgaa
     1981 cagatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     2041 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     2101 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     2161 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     2221 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     2281 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     2341 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     2401 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     2461 tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac
     2521 ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg
     2581 ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcggcg agcggtatca
     2641 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2701 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2761 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2821 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2881 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2941 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3001 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3061 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3121 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3181 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3241 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3301 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3361 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3421 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3481 tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg
     3541 gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg
     3601 cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac
     3661 cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt
     3721 gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc
     3781 ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     3841 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     3901 cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga
     3961 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     4021 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     4081 ccaacgctat gtcctgatag cgatccgcca cacccagccg gccacagtcg atgaatccag
     4141 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     4201 gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     4261 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     4321 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     4381 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     4441 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     4501 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     4561 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     4621 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     4681 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     4741 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg
     4801 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     4861 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     4921 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     4981 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     5041 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     5101 cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc ctttttcaat
     5161 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5221 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
     5281 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5341 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5401 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5461 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5521 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5581 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5641 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5701 attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg
     5761 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5821 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     5881 ggccagtgcc aagct
//
