LOCUS       sequence-222237-        5922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of the human NODAL 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 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: NODAL-MYC-DYK-C312A
FEATURES             Location/Qualifiers
     source          1..5922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        108..487
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        488..691
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     642..662
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     717..736
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        717..735
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1853..1882
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1901..1924
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1944..1960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1944..1960)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1971..2593
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2170..2189)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2742..3330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2822..2841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3659..3706
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3731..3750
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3938..4732)
                     /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(3938..4732)
                     /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(4049..4068)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4659..4678
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4767..5124)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4781..4916
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4933..5124
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5081..5101
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5126..5230)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5257..5712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5344..5363)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5554..5575
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5838..5860
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5852..5869
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5853..5869
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tggttatata gcataaatca atattggcta ttggccattg catacgttgt atctatatca
       61 taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt
      121 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      181 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      241 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      301 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      361 gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      421 gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      481 taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg
      541 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      601 acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg
      661 tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga attttgtaat
      721 acgactcact atagggcggc cgggaattcg tcgactggat ccggtaccga ggagatctgc
      781 cgccgcgatc gccatgcacg cccactgcct gcccttcctt ctgcacgcct ggtgggccct
      841 actccaggcg ggtgctgcga cggtggccac tgcgctcctg cgtacgcggg ggcagccctc
      901 gtcgccatcc cctctggcgt acatgctgag cctctaccgc gacccgctgc cgagggcaga
      961 catcatccgc agcctacagg cagaagatgt ggcagtggat gggcagaact ggacgtttgc
     1021 ttttgacttc tccttcctga gccaacaaga ggatctggca tgggctgagc tccggctgca
     1081 gctgtccagc cctgtggacc tccccactga gggctcactt gccattgaga ttttccacca
     1141 gccaaagccc gacacagagc aggcttcaga cagctgctta gagcggtttc agatggacct
     1201 attcactgtc actttgtccc aggtcacctt ttccttgggc agcatggttt tggaggtgac
     1261 caggcctctc tccaagtggc tgaagcgccc tggggccctg gagaagcaga tgtccagggt
     1321 agctggagag tgctggccgc ggccccccac accgcctgcc accaatgtgc tccttatgct
     1381 ctactccaac ctctcgcagg agcagaggca gctgggtggg tccaccttgc tgtgggaagc
     1441 cgagagctcc tggcgggccc aggagggaca gctgtcctgg gagtggggca agaggcaccg
     1501 tcgacatcac ttgccagaca gaagtcaact gtgtcggaag gtcaagttcc aggtggactt
     1561 caacctgatc ggatggggct cctggatcat ctaccccaag cagtacaacg cctatcgctg
     1621 tgagggcgag tgtcctaatc ctgttgggga ggagtttcat ccgaccaacc atgcatacat
     1681 ccagagtctg ctgaaacgtt accagcccca ccgagtccct tccactgcct gtgccccagt
     1741 gaagaccaag ccgctgagca tgctgtatgt ggataatggc agagtgctcc tagatcacca
     1801 taaagacatg atcgtggaag aatgtgggtg cctcacgcgt acgcggccgc tcgagcagaa
     1861 actcatctca gaagaggatc tggcagcaaa tgatatcctg gattacaagg atgacgacga
     1921 taaggtttaa acggccggcc gcggtcatag ctgtttcctg aacagatccc gggtggcatc
     1981 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     2041 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     2101 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     2161 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     2221 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     2281 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     2341 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     2401 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
     2461 aactatacca gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg
     2521 tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga
     2581 tgcctgttga attgggtacg cggccagcgg cgagcggtat cagctcactc aaaggcggta
     2641 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2701 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2761 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2821 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2881 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2941 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3001 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3061 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3121 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3181 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3241 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3301 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3361 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3421 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3481 gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct
     3541 tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa
     3601 tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga
     3661 acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt
     3721 ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac
     3781 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     3841 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     3901 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     3961 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     4021 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     4081 agcgatccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     4141 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     4201 tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     4261 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     4321 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     4381 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     4441 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     4501 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat
     4561 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     4621 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     4681 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     4741 atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa aaagcctcct
     4801 cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa
     4861 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     4921 ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg
     4981 cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt
     5041 tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc
     5101 cacagctggt tctttccgcc tcaggactct tcctttttca atattattga agcatttatc
     5161 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5221 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     5281 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5341 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5401 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5461 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5521 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5581 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5641 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5701 cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     5761 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     5821 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctgat
     5881 ctatacattg aatcaatatt ggcaattagc catattagtc at
//
