LOCUS       sequence-222245-        5895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of a human NODAL splice variant open 
            reading frame (with a mutated Cysteine residue) 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-C302A
FEATURES             Location/Qualifiers
     source          1..5895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        158..537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        538..741
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     692..712
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     767..786
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        767..785
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1876..1905
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1924..1947
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1967..1983)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1967..1983)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1994..2616
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2193..2212)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2765..3353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2845..2864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3682..3729
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3754..3773
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3961..4755)
                     /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(3961..4755)
                     /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(4072..4091)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4682..4701
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4790..5147)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4804..4939
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4956..5147
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5104..5124
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5149..5253)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5280..5735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5367..5386)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5577..5598
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5861..5883
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5875..5892
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5876..5892
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagctgatct atacattgaa tcaatattgg caattagcca tattagtcat tggttatata
       61 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
      121 atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt gactagttat
      181 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      241 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      301 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      361 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg
      421 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      481 ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      541 atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca
      601 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      661 ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg
      721 gaggtctata taagcagagc tcgtttagtg aaccgtcaga attttgtaat acgactcact
      781 atagggcggc cgggaattcg tcgactggat ccggtaccga ggagatctgc cgccgcgatc
      841 gccatgcacg cccactgcct gcccttcctt ctgcacgcct ggtgggccct actccaggcg
      901 ggtgctgcga cggtggccac tgcgctcctg cgtacgcggg ggcagccctc gtcgccatcc
      961 cctctggcgt acatgctgag cctctaccgc gacccgctgc cgagggcaga catcatccgc
     1021 agcctacagg cagaagatgt ggcagtggat gggcagaact ggacgtttgc ttttgacttc
     1081 tccttcctga gccaacaaga ggatctggca tgggctgagc tccggctgca gctgtccagc
     1141 cctgtggacc tccccactga gggctcactt gccattgaga ttttccacca gccaaagccc
     1201 gacacagagc aggcttcaga cagctgctta gagcggtttc agatggacct attcactgtc
     1261 actttgtccc aggtcacctt ttccttgggc agcatggttt tggaggtgac caggcctctc
     1321 tccaagtggc tgaagcaccc tggggccctg gagaagcaga tgtccagggt agctggagag
     1381 tgctggccgc ggccccccac accgcctgcc accaatgtgc tccttatgct ctactccaac
     1441 ctctcgcagg agcagaggca gctgggtggg tccaccttgc tgtgggaagc cgagagctcc
     1501 tggcgggccc aggagggaca gctgtcctgg gagtggggca agaggcaccg tcgacatcac
     1561 ttgccagaca gaagtcaact gtgtcggaag gtcaagttcc aggtggactt caacctgatc
     1621 ggatggggct cctggatcat ctaccccaag cagtacaacg cctatcgctg tgagggcgag
     1681 tgtcctaatc ctgttgggga ggagtttcat ccgaccaacc atgcatacat ccaggtggcc
     1741 ctgcccgcct gtccaaggtc atatgggacc aaaatgtttt cattttactc catgaagtct
     1801 ggaatgagaa tttcttggac ttgcaatata tcctccatgc caagcctcag agtctgcacg
     1861 cgtacgcggc cgctcgagca gaaactcatc tcagaagagg atctggcagc aaatgatatc
     1921 ctggattaca aggatgacga cgataaggtt taaacggccg gccgcggtca tagctgtttc
     1981 ctgaacagat cccgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     2041 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     2101 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     2161 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     2221 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     2281 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     2341 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     2401 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt
     2461 ccctgtcctt ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag
     2521 gctacctggc catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc
     2581 atgcgttggg tccactcagt agatgcctgt tgaattgggt acgcggccag cggcgagcgg
     2641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2701 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2941 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3181 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3301 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3361 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3421 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3481 ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga
     3541 cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag
     3601 aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct
     3661 gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt
     3721 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc
     3781 tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     3841 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     3901 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     3961 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     4021 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     4081 ggtagccaac gctatgtcct gatagcgatc cgccacaccc agccggccac agtcgatgaa
     4141 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     4201 gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc
     4261 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     4321 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     4381 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     4441 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     4501 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     4561 cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     4621 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     4681 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     4741 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct ttgcaaaagc
     4801 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     4861 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     4921 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     4981 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     5041 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     5101 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcaggac tcttcctttt
     5161 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5221 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5281 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5341 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5401 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5461 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5521 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5581 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5641 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5701 cgcgaatttt aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac
     5761 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     5821 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     5881 acgacggcca gtgcc
//
