LOCUS       sequence-222166-        5895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of a human NODAL splice variant open 
            reading frame (with a mutated N-glycosylation site) 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-N328A
FEATURES             Location/Qualifiers
     source          1..5895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..19
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3..19
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        180..559
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        560..763
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     714..734
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     789..808
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        789..807
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1898..1927
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1946..1969
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1989..2005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1989..2005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2016..2638
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2215..2234)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2787..3375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2867..2886)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3704..3751
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3776..3795
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3983..4777)
                     /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(3983..4777)
                     /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(4094..4113)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4704..4723
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4812..5169)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4826..4961
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4978..5169
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5126..5146
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5171..5275)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5302..5757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5389..5408)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5599..5620
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(5883..5895,1..10)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgtaaaacg acggccagtg ccaagctgat ctatacattg aatcaatatt ggcaattagc
       61 catattagtc attggttata tagcataaat caatattggc tattggccat tgcatacgtt
      121 gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg
      181 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      241 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      301 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      361 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      421 agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      481 gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt
      541 agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg
      601 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      661 gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat
      721 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
      781 gaattttgta atacgactca ctatagggcg gccgggaatt cgtcgactgg atccggtacc
      841 gaggagatct gccgccgcga tcgccatgca cgcccactgc ctgcccttcc ttctgcacgc
      901 ctggtgggcc ctactccagg cgggtgctgc gacggtggcc actgcgctcc tgcgtacgcg
      961 ggggcagccc tcgtcgccat cccctctggc gtacatgctg agcctctacc gcgacccgct
     1021 gccgagggca gacatcatcc gcagcctaca ggcagaagat gtggcagtgg atgggcagaa
     1081 ctggacgttt gcttttgact tctccttcct gagccaacaa gaggatctgg catgggctga
     1141 gctccggctg cagctgtcca gccctgtgga cctccccact gagggctcac ttgccattga
     1201 gattttccac cagccaaagc ccgacacaga gcaggcttca gacagctgct tagagcggtt
     1261 tcagatggac ctattcactg tcactttgtc ccaggtcacc ttttccttgg gcagcatggt
     1321 tttggaggtg accaggcctc tctccaagtg gctgaagcac cctggggccc tggagaagca
     1381 gatgtccagg gtagctggag agtgctggcc gcggcccccc acaccgcctg ccaccaatgt
     1441 gctccttatg ctctactcca acctctcgca ggagcagagg cagctgggtg ggtccacctt
     1501 gctgtgggaa gccgagagct cctggcgggc ccaggaggga cagctgtcct gggagtgggg
     1561 caagaggcac cgtcgacatc acttgccaga cagaagtcaa ctgtgtcgga aggtcaagtt
     1621 ccaggtggac ttcaacctga tcggatgggg ctcctggatc atctacccca agcagtacaa
     1681 cgcctatcgc tgtgagggcg agtgtcctaa tcctgttggg gaggagtttc atccgaccaa
     1741 ccatgcatac atccaggtgg ccctgccctg ctgtccaagg tcatatggga ccaaaatgtt
     1801 ttcattttac tccatgaagt ctggaatgag aatttcttgg acttgcgcta tatcctccat
     1861 gccaagcctc agagtctgca cgcgtacgcg gccgctcgag cagaaactca tctcagaaga
     1921 ggatctggca gcaaatgata tcctggatta caaggatgac gacgataagg tttaaacggc
     1981 cggccgcggt catagctgtt tcctgaacag atcccgggtg gcatccctgt gacccctccc
     2041 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     2101 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     2161 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     2221 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     2281 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     2341 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     2401 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     2461 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg
     2521 aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga catttgagtt
     2581 gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg
     2641 gtacgcggcc agcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2701 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2761 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2821 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2881 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2941 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3001 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3061 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3121 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3181 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3241 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3301 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3361 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3421 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3481 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct
     3541 atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg
     3601 gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg
     3661 gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca
     3721 ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc
     3781 tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc
     3841 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     3901 tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc
     3961 atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc
     4021 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     4081 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcga tccgccacac
     4141 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     4201 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     4261 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     4321 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     4381 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     4441 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     4501 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     4561 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     4621 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     4681 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     4741 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     4801 cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     4861 agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg
     4921 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     4981 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     5041 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     5101 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
     5161 ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5221 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5281 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5341 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5401 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5461 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5521 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5581 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5641 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5701 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca
     5761 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     5821 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     5881 ttcccagtca cgacg
//
