LOCUS       sequence-222486-        5949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of the human NODAL open reading frame with 
            an internal DYK tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5949)
  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 5949)
  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-matDYK
FEATURES             Location/Qualifiers
     source          1..5949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..37
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     29..46
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     30..46
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        207..586
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        587..790
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     741..761
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     816..835
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        816..834
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1607..1630
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1979..2008
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2027..2050
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2070..2086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2070..2086)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2097..2719
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2296..2315)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2868..3456)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2948..2967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3785..3832
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3857..3876
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4064..4858)
                     /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(4064..4858)
                     /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(4175..4194)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4785..4804
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4893..5250)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4907..5042
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5059..5250
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5207..5227
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5252..5356)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5383..5838
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5470..5489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5680..5701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctgatcta
       61 tacattgaat caatattggc aattagccat attagtcatt ggttatatag cataaatcaa
      121 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
      181 ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc
      241 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      301 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      361 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      421 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
      481 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
      541 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      601 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      661 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      721 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      781 aagcagagct cgtttagtga accgtcagaa ttttgtaata cgactcacta tagggcggcc
      841 gggaattcat cgactggatc cggtaccgag gagatctgcc gccgcgatcg ccatgcacgc
      901 ccactgcctg cccttccttc tgcacgcctg gtgggcccta ctccaggcgg gtgctgcgac
      961 ggtggccact gcgctcctgc gtacgcgggg gcagccctcg tcgccatccc ctctggcgta
     1021 catgctgagc ctctaccgcg acccgctgcc gagggcagac atcatccgca gcctacaggc
     1081 agaagatgtg gcagtggatg ggcagaactg gacgtttgct tttgacttct ccttcctgag
     1141 ccaacaagag gatctggcat gggctgagct ccggctgcag ctgtccagcc ctgtggacct
     1201 ccccactgag ggctcacttg ccattgagat tttccaccag ccaaagcccg acacagagca
     1261 ggcttcagac agctgcttag agcggtttca gatggaccta ttcactgtca ctttgtccca
     1321 ggtcaccttt tccttgggca gcatggtttt ggaggtgacc aggcctctct ccaagtggct
     1381 gaagcaccct ggggccctgg agaagcagat gtccagggta gctggagagt gctggccgcg
     1441 gccccccaca ccgcctgcca ccaatgtgct ccttatgctc tactccaacc tctcgcagga
     1501 gcagaggcag ctgggtgggt ccaccttgct gtgggaagcc gagagctcct ggcgggccca
     1561 ggagggacag ctgtcctggg agtggggcaa gaggcaccgt cgacatgatt acaaggatga
     1621 cgacgataag catcacttgc cagacagaag tcaactgtgt cggaaggtca agttccaggt
     1681 ggacttcaac ctgatcggat ggggctcctg gatcatctac cccaagcagt acaacgccta
     1741 tcgctgtgag ggcgagtgtc ctaatcctgt tggggaggag tttcatccga ccaaccatgc
     1801 atacatccag agtctgctga aacgttacca gccccaccga gtcccttcca cttgttgtgc
     1861 cccagtgaag accaagccgc tgagcatgct gtatgtggat aatggcagag tgctcctaga
     1921 tcaccataaa gacatgatcg tggaagaatg tgggtgcctc tgacgtacgc ggccgctcga
     1981 gcagaaactc atctcagaag aggatctggc agcaaatgat atcctggatt acaaggatga
     2041 cgacgataag gtttaaacgg ccggccgcgg tcatagctgt ttcctgaaca gatcccgggt
     2101 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     2161 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     2221 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     2281 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     2341 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     2401 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     2461 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     2521 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     2581 taaaataact ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc
     2641 aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc
     2701 agtagatgcc tgttgaattg ggtacgcggc cagcggcgag cggtatcagc tcactcaaag
     2761 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2821 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2881 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2941 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3001 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3061 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3121 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3181 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3241 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3301 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3361 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3421 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3481 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3541 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3601 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
     3661 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
     3721 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     3781 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     3841 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
     3901 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     3961 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     4021 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     4081 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     4141 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     4201 cctgatagcg atccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     4261 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     4321 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     4381 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     4441 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     4501 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     4561 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     4621 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
     4681 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     4741 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     4801 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     4861 gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag
     4921 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     4981 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     5041 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     5101 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     5161 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     5221 acattccaca gctggttctt tccgcctcag gactcttcct ttttcaatat tattgaagca
     5281 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5341 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     5401 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5461 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5521 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5581 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5641 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5701 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5761 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5821 tattaacgct tacaatttcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5881 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5941 aagttgggt
//
