LOCUS       sequence-222469-        5922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of a human NODAL splice varaint open 
            reading frame with an internal DYK tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5922)
  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 5922)
  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-matDYK
FEATURES             Location/Qualifiers
     source          1..5922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        94..473
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        474..677
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     628..648
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     703..722
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        703..721
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1494..1517
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1839..1868
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1887..1910
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1930..1946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1930..1946)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1957..2579
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2156..2175)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2728..3316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2808..2827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3645..3692
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3717..3736
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3924..4718)
                     /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(3924..4718)
                     /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(4035..4054)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4645..4664
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4753..5110)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4767..4902
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4919..5110
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5067..5087
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5112..5216)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5243..5698
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5330..5349)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5540..5561
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5824..5846
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5838..5855
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5839..5855
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
       61 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat
      121 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      181 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      241 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      301 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
      361 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
      421 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      481 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      541 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      601 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
      661 tctatataag cagagctcgt ttagtgaacc gtcagaattt tgtaatacga ctcactatag
      721 ggcggccggg aattcatcga ctggatccgg taccgaggag atctgccgcc gcgatcgcca
      781 tgcacgccca ctgcctgccc ttccttctgc acgcctggtg ggccctactc caggcgggtg
      841 ctgcgacggt ggccactgcg ctcctgcgta cgcgggggca gccctcgtcg ccatcccctc
      901 tggcgtacat gctgagcctc taccgcgacc cgctgccgag ggcagacatc atccgcagcc
      961 tacaggcaga agatgtggca gtggatgggc agaactggac gtttgctttt gacttctcct
     1021 tcctgagcca acaagaggat ctggcatggg ctgagctccg gctgcagctg tccagccctg
     1081 tggacctccc cactgagggc tcacttgcca ttgagatttt ccaccagcca aagcccgaca
     1141 cagagcaggc ttcagacagc tgcttagagc ggtttcagat ggacctattc actgtcactt
     1201 tgtcccaggt caccttttcc ttgggcagca tggttttgga ggtgaccagg cctctctcca
     1261 agtggctgaa gcaccctggg gccctggaga agcagatgtc cagggtagct ggagagtgct
     1321 ggccgcggcc ccccacaccg cctgccacca atgtgctcct tatgctctac tccaacctct
     1381 cgcaggagca gaggcagctg ggtgggtcca ccttgctgtg ggaagccgag agctcctggc
     1441 gggcccagga gggacagctg tcctgggagt ggggcaagag gcaccgtcga catgattaca
     1501 aggatgacga cgataagcat cacttgccag acagaagtca actgtgtcgg aaggtcaagt
     1561 tccaggtgga cttcaacctg atcggatggg gctcctggat catctacccc aagcagtaca
     1621 acgcctatcg ctgtgagggc gagtgtccta atcctgttgg ggaggagttt catccgacca
     1681 accatgcata catccaggtg gccctgccct gctgtccaag gtcatatggg accaaaatgt
     1741 tttcatttta ctccatgaag tctggaatga gaatttcttg gacttgcaat atatcctcca
     1801 tgccaagcct cagagtctgc tgacgtacgc ggccgctcga gcagaaactc atctcagaag
     1861 aggatctggc agcaaatgat atcctggatt acaaggatga cgacgataag gtttaaacgg
     1921 ccggccgcgg tcatagctgt ttcctgaaca gatcccgggt ggcatccctg tgacccctcc
     1981 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     2041 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     2101 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     2161 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     2221 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     2281 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     2341 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     2401 caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag
     2461 gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt
     2521 tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg
     2581 ggtacgcggc cagcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2641 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2701 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2761 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2821 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2881 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2941 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3001 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3061 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3121 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3181 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3241 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3301 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3361 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3421 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aacctgaggc
     3481 tatggcaggg cctgccgccc cgacgttggc tgcgagccct gggccttcac ccgaacttgg
     3541 ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg
     3601 ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa acgacccaac
     3661 accgtgcgtt ttattctgtc tttttattgc cgtcatagcg cgggttcctt ccggtattgt
     3721 ctccttccgt gtttcagtta gcctccccct agggtgggcg aagaactcca gcatgagatc
     3781 cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc
     3841 atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt
     3901 catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg
     3961 cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
     4021 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg atccgccaca
     4081 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
     4141 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgct cgccttgagc
     4201 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
     4261 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
     4321 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
     4381 actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat
     4441 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
     4501 gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac
     4561 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
     4621 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
     4681 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc
     4741 tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     4801 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     4861 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     4921 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     4981 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     5041 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     5101 tccgcctcag gactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5161 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5221 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5281 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5341 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5401 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5461 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5521 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5581 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5641 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     5701 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5761 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     5821 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc
     5881 aatattggca attagccata ttagtcattg gttatatagc at
//
