LOCUS       sequence-222487-        5991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of a C-terminal truncated human NODAL open 
            reading frame with an internal MYC tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5991)
  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 5991)
  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-matMYC-ex2trunc
FEATURES             Location/Qualifiers
     source          1..5991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..459
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        460..663
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     614..634
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     689..708
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        689..707
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1480..1509
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1894..1923
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1942..1965
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1985..2001)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1985..2001)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2012..2634
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2211..2230)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2783..3371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2863..2882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3700..3747
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3772..3791
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3979..4773)
                     /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(3979..4773)
                     /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(4090..4109)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4700..4719
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4808..5165)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4822..4957
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4974..5165
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5122..5142
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5167..5271)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5298..5753
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5385..5404)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5595..5616
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5879..5901
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5893..5910
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5894..5910
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt
       61 ccaatatgac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg
      121 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      181 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      241 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      301 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
      361 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
      421 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      481 accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      541 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac
      601 cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      661 gctcgtttag tgaaccgtca gaattttgta atacgactca ctatagggcg gccgggaatt
      721 catcgactgg atccggtacc gaggagatct gccgccgcga tcgccatgca cgcccactgc
      781 ctgcccttcc ttctgcacgc ctggtgggcc ctactccagg cgggtgctgc gacggtggcc
      841 actgcgctcc tgcgtacgcg ggggcagccc tcgtcgccat cccctctggc gtacatgctg
      901 agcctctacc gcgacccgct gccgagggca gacatcatcc gcagcctaca ggcagaagat
      961 gtggcagtgg atgggcagaa ctggacgttt gcttttgact tctccttcct gagccaacaa
     1021 gaggatctgg catgggctga gctccggctg cagctgtcca gccctgtgga cctccccact
     1081 gagggctcac ttgccattga gattttccac cagccaaagc ccgacacaga gcaggcttca
     1141 gacagctgct tagagcggtt tcagatggac ctattcactg tcactttgtc ccaggtcacc
     1201 ttttccttgg gcagcatggt tttggaggtg accaggcctc tctccaagtg gctgaagcac
     1261 cctggggccc tggagaagca gatgtccagg gtagctggag agtgctggcc gcggcccccc
     1321 acaccgcctg ccaccaatgt gctccttatg ctctactcca acctctcgca ggagcagagg
     1381 cagctgggtg ggtccacctt gctgtgggaa gccgagagct cctggcgggc ccaggaggga
     1441 cagctgtcct gggagtgggg caagaggcac cgtcgacatg agcagaaact catctcagaa
     1501 gaggatctgc atcacttgcc agacagaagt caactgtgtc ggaaggtcaa gttccaggtg
     1561 gacttcaacc tgatcggatg gggctcctgg atcatctacc ccaagcagta caacgcctat
     1621 cgctgtgagg gcgagtgtcc taatcctgtt ggggaggagt ttcatccgac caaccatgca
     1681 tacatccagt gactgctgaa acgttaccag catgcataca tccagtgact gctgaaacgt
     1741 taccagcccc accgagtccc ttccacttgt tgtgccccag tgaagaccaa gccgctgagc
     1801 atgctgtatg tggataatgg cagagtgctc ctagatcacc ataaagacat gatcgtggaa
     1861 gaatgtgggt gcctctgacg tacgcggccg ctcgagcaga aactcatctc agaagaggat
     1921 ctggcagcaa atgatatcct ggattacaag gatgacgacg ataaggttta aacggccggc
     1981 cgcggtcata gctgtttcct gaacagatcc cgggtggcat ccctgtgacc cctccccagt
     2041 gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt
     2101 aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt
     2161 ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga
     2221 accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca
     2281 agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct
     2341 cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc
     2401 caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc
     2461 gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga
     2521 cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt
     2581 gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac
     2641 gcggccagcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2701 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2761 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2821 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2881 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2941 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3001 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3061 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3121 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3181 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3241 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3301 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3361 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3421 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3481 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg
     3541 cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt
     3601 ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat
     3661 cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt
     3721 gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct
     3781 tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc
     3841 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     3901 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     3961 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     4021 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     4081 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcgatccg ccacacccag
     4141 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     4201 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc
     4261 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     4321 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     4381 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     4441 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     4501 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     4561 ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc
     4621 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     4681 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     4741 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     4801 atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     4861 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     4921 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     4981 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     5041 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5101 gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc
     5161 ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5221 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5281 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5341 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5401 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5461 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5521 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5581 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5641 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5701 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttccattcg
     5761 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     5821 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     5881 cagtcacgac gttgtaaaac gacggccagt gccaagctga tctatacatt gaatcaatat
     5941 tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg c
//
