LOCUS       sequence-224221-        5955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of the 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 5955)
  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 5955)
  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
FEATURES             Location/Qualifiers
     source          1..5955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..54
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     46..63
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     47..63
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        224..603
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        604..807
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     758..778
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     833..852
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        833..851
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1624..1653
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2002..2031
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2050..2073
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2093..2109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2093..2109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2120..2742
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2319..2338)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2891..3479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2971..2990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3808..3855
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3880..3899
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4087..4881)
                     /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(4087..4881)
                     /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(4198..4217)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4808..4827
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4916..5273)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4930..5065
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5082..5273
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5230..5250
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5275..5379)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5406..5861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5493..5512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5703..5724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
       61 agtgccaagc tgatctatac attgaatcaa tattggcaat tagccatatt agtcattggt
      121 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
      181 atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg attattgact
      241 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      301 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      361 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      421 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      481 agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      541 atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      601 atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga
      661 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      721 gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta
      781 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagaattt tgtaatacga
      841 ctcactatag ggcggccggg aattcatcga ctggatccgg taccgaggag atctgccgcc
      901 gcgatcgcca tgcacgccca ctgcctgccc ttccttctgc acgcctggtg ggccctactc
      961 caggcgggtg ctgcgacggt ggccactgcg ctcctgcgta cgcgggggca gccctcgtcg
     1021 ccatcccctc tggcgtacat gctgagcctc taccgcgacc cgctgccgag ggcagacatc
     1081 atccgcagcc tacaggcaga agatgtggca gtggatgggc agaactggac gtttgctttt
     1141 gacttctcct tcctgagcca acaagaggat ctggcatggg ctgagctccg gctgcagctg
     1201 tccagccctg tggacctccc cactgagggc tcacttgcca ttgagatttt ccaccagcca
     1261 aagcccgaca cagagcaggc ttcagacagc tgcttagagc ggtttcagat ggacctattc
     1321 actgtcactt tgtcccaggt caccttttcc ttgggcagca tggttttgga ggtgaccagg
     1381 cctctctcca agtggctgaa gcaccctggg gccctggaga agcagatgtc cagggtagct
     1441 ggagagtgct ggccgcggcc ccccacaccg cctgccacca atgtgctcct tatgctctac
     1501 tccaacctct cgcaggagca gaggcagctg ggtgggtcca ccttgctgtg ggaagccgag
     1561 agctcctggc gggcccagga gggacagctg tcctgggagt ggggcaagag gcaccgtcga
     1621 catgagcaga aactcatctc agaagaggat ctgcatcact tgccagacag aagtcaactg
     1681 tgtcggaagg tcaagttcca ggtggacttc aacctgatcg gatggggctc ctggatcatc
     1741 taccccaagc agtacaacgc ctatcgctgt gagggcgagt gtcctaatcc tgttggggag
     1801 gagtttcatc cgaccaacca tgcatacatc cagagtctgc tgaaacgtta ccagccccac
     1861 cgagtccctt ccacttgttg tgccccagtg aagaccaagc cgctgagcat gctgtatgtg
     1921 gataatggca gagtgctcct agatcaccat aaagacatga tcgtggaaga atgtgggtgc
     1981 ctctgacgta cgcggccgct cgagcagaaa ctcatctcag aagaggatct ggcagcaaat
     2041 gatatcctgg attacaagga tgacgacgat aaggtttaaa cggccggccg cggtcatagc
     2101 tgtttcctga acagatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     2161 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     2221 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     2281 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     2341 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     2401 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     2461 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     2521 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     2581 tcccttccct gtccttctga ttttaaaata actataccag caggaggacg tccagacaca
     2641 gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt
     2701 cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcggc
     2761 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2821 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2881 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2941 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3001 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3061 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3121 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3181 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3241 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3301 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3361 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3421 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3481 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3541 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3601 gaagttttaa atcaatctaa agtatatatg agtaacctga ggctatggca gggcctgccg
     3661 ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa
     3721 aggaagaaac gcgggcgtat tggccccaat ggggtctcgg tggggtatcg acagagtgcc
     3781 agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct
     3841 gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag
     3901 ttagcctccc cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     3961 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     4021 aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag
     4081 tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     4141 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
     4201 atcacgggta gccaacgcta tgtcctgata gcgatccgcc acacccagcc ggccacagtc
     4261 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
     4321 ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc
     4381 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
     4441 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     4501 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     4561 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     4621 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     4681 tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     4741 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     4801 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     4861 caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cgatctttgc
     4921 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     4981 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     5041 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     5101 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     5161 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     5221 actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct caggactctt
     5281 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5341 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5401 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5461 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5521 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5581 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5641 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5701 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5761 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5821 atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg
     5881 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     5941 gggggatgtg ctgca
//
