LOCUS       sequence-217102-        5955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of the human NODAL open reading frame with 
            an internal MYC tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5955)
  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 5955)
  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-matMYC
FEATURES             Location/Qualifiers
     source          1..5955
                     /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..1772
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2121..2150
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2169..2192
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2212..2228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2212..2228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2239..2861
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2438..2457)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(3010..3598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3090..3109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3927..3974
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3999..4018
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4206..5000)
                     /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(4206..5000)
                     /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(4317..4336)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4927..4946
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5035..5392)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5049..5184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5201..5392
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5349..5369
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5394..5498)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5525..5953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5612..5631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5822..5843
                     /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 gcaggagcag aggcagctgg gtgggtccac cttgctgtgg gaagccgaga
     1681 gctcctggcg ggcccaggag ggacagctgt cctgggagtg gggcaagagg caccgtcgac
     1741 atgagcagaa actcatctca gaagaggatc tgcatcactt gccagacaga agtcaactgt
     1801 gtcggaaggt caagttccag gtggacttca acctgatcgg atggggctcc tggatcatct
     1861 accccaagca gtacaacgcc tatcgctgtg agggcgagtg tcctaatcct gttggggagg
     1921 agtttcatcc gaccaaccat gcatacatcc agagtctgct gaaacgttac cagccccacc
     1981 gagtcccttc cacttgttgt gccccagtga agaccaagcc gctgagcatg ctgtatgtgg
     2041 ataatggcag agtgctccta gatcaccata aagacatgat cgtggaagaa tgtgggtgcc
     2101 tctgacgtac gcggccgctc gagcagaaac tcatctcaga agaggatctg gcagcaaatg
     2161 atatcctgga ttacaaggat gacgacgata aggtttaaac ggccggccgc ggtcatagct
     2221 gtttcctgaa cagatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     2281 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     2341 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     2401 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     2461 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     2521 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     2581 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     2641 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     2701 cccttccctg tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag
     2761 cataggctac ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc
     2821 ctctcatgcg ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcggcg
     2881 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2941 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3421 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3541 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3721 aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc
     3781 cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa
     3841 ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca
     3901 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg
     3961 tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt
     4021 tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
     4081 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
     4141 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     4201 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     4261 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     4321 tcacgggtag ccaacgctat gtcctgatag cgatccgcca cacccagccg gccacagtcg
     4381 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     4441 gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct
     4501 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     4561 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     4621 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     4681 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     4741 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     4801 agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     4861 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     4921 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     4981 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca
     5041 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     5101 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     5161 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     5221 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     5281 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5341 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc
     5401 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5461 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5521 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5581 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5641 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5701 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5761 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5821 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5881 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5941 tttaacgcga atttt
//
