LOCUS       sequence-222161-        5895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of a human NODAL splice variant open 
            reading frame 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
FEATURES             Location/Qualifiers
     source          1..5895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        161..540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        541..744
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     695..715
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     770..789
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        770..788
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1879..1908
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1927..1950
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1970..1986)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1970..1986)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1997..2619
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2196..2215)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2768..3356)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2848..2867)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3685..3732
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3757..3776
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3964..4758)
                     /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(3964..4758)
                     /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(4075..4094)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4685..4704
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4793..5150)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4807..4942
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4959..5150
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5107..5127
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5152..5256)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5283..5738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5370..5389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5580..5601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5864..5886
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5878..5895
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5879..5895
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gccaagctga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat
       61 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
      121 tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt
      181 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      241 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      301 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      361 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      421 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      481 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      541 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
      601 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      661 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      721 tgggaggtct atataagcag agctcgttta gtgaaccgtc agaattttgt aatacgactc
      781 actatagggc ggccgggaat tcgtcgactg gatccggtac cgaggagatc tgccgccgcg
      841 atcgccatgc acgcccactg cctgcccttc cttctgcacg cctggtgggc cctactccag
      901 gcgggtgctg cgacggtggc cactgcgctc ctgcgtacgc gggggcagcc ctcgtcgcca
      961 tcccctctgg cgtacatgct gagcctctac cgcgacccgc tgccgagggc agacatcatc
     1021 cgcagcctac aggcagaaga tgtggcagtg gatgggcaga actggacgtt tgcttttgac
     1081 ttctccttcc tgagccaaca agaggatctg gcatgggctg agctccggct gcagctgtcc
     1141 agccctgtgg acctccccac tgagggctca cttgccattg agattttcca ccagccaaag
     1201 cccgacacag agcaggcttc agacagctgc ttagagcggt ttcagatgga cctattcact
     1261 gtcactttgt cccaggtcac cttttccttg ggcagcatgg ttttggaggt gaccaggcct
     1321 ctctccaagt ggctgaagca ccctggggcc ctggagaagc agatgtccag ggtagctgga
     1381 gagtgctggc cgcggccccc cacaccgcct gccaccaatg tgctccttat gctctactcc
     1441 aacctctcgc aggagcagag gcagctgggt gggtccacct tgctgtggga agccgagagc
     1501 tcctggcggg cccaggaggg acagctgtcc tgggagtggg gcaagaggca ccgtcgacat
     1561 cacttgccag acagaagtca actgtgtcgg aaggtcaagt tccaggtgga cttcaacctg
     1621 atcggatggg gctcctggat catctacccc aagcagtaca acgcctatcg ctgtgagggc
     1681 gagtgtccta atcctgttgg ggaggagttt catccgacca accatgcata catccaggtg
     1741 gccctgccct gctgtccaag gtcatatggg accaaaatgt tttcatttta ctccatgaag
     1801 tctggaatga gaatttcttg gacttgcaat atatcctcca tgccaagcct cagagtctgc
     1861 acgcgtacgc ggccgctcga gcagaaactc atctcagaag aggatctggc agcaaatgat
     1921 atcctggatt acaaggatga cgacgataag gtttaaacgg ccggccgcgg tcatagctgt
     1981 ttcctgaaca gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     2041 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     2101 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     2161 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     2221 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     2281 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     2341 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     2401 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     2461 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca
     2521 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct
     2581 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcggcgag
     2641 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2701 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2761 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2821 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2881 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2941 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3001 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3061 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3121 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3181 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3241 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3301 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3361 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3421 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3481 gttttaaatc aatctaaagt atatatgagt aacctgaggc tatggcaggg cctgccgccc
     3541 cgacgttggc tgcgagccct gggccttcac ccgaacttgg ggggtggggt ggggaaaagg
     3601 aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc
     3661 cctgggaccg aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt ttattctgtc
     3721 tttttattgc cgtcatagcg cgggttcctt ccggtattgt ctccttccgt gtttcagtta
     3781 gcctccccct agggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     3841 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     3901 cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc
     3961 cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     4021 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     4081 acgggtagcc aacgctatgt cctgatagcg atccgccaca cccagccggc cacagtcgat
     4141 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     4201 cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg
     4261 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     4321 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     4381 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     4441 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     4501 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     4561 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     4621 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     4681 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     4741 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa
     4801 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     4861 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     4921 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     4981 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     5041 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     5101 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag gactcttcct
     5161 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5221 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5281 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5341 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5401 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5461 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5521 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5581 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5641 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5701 taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc
     5761 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     5821 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     5881 aaaacgacgg ccagt
//
