LOCUS       sequence-222231-        5928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of a human NODAL splice variant open 
            reading frame with an internal MYC tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5928)
  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 5928)
  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-matMYC
FEATURES             Location/Qualifiers
     source          1..5928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        85..464
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        465..668
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     619..639
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     694..713
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        694..712
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1485..1514
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1836..1865
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1884..1907
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1927..1943)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1927..1943)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1954..2576
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2153..2172)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2725..3313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2805..2824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3642..3689
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3714..3733
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3921..4715)
                     /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(3921..4715)
                     /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(4032..4051)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4642..4661
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4750..5107)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4764..4899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4916..5107
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5064..5084
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5109..5213)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5240..5695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5327..5346)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5537..5558
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5821..5843
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5835..5852
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5836..5852
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
       61 catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
      121 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      181 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      241 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      301 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg
      361 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc
      421 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      481 agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      541 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      601 ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      661 gcagagctcg tttagtgaac cgtcagaatt ttgtaatacg actcactata gggcggccgg
      721 gaattcatcg actggatccg gtaccgagga gatctgccgc cgcgatcgcc atgcacgccc
      781 actgcctgcc cttccttctg cacgcctggt gggccctact ccaggcgggt gctgcgacgg
      841 tggccactgc gctcctgcgt acgcgggggc agccctcgtc gccatcccct ctggcgtaca
      901 tgctgagcct ctaccgcgac ccgctgccga gggcagacat catccgcagc ctacaggcag
      961 aagatgtggc agtggatggg cagaactgga cgtttgcttt tgacttctcc ttcctgagcc
     1021 aacaagagga tctggcatgg gctgagctcc ggctgcagct gtccagccct gtggacctcc
     1081 ccactgaggg ctcacttgcc attgagattt tccaccagcc aaagcccgac acagagcagg
     1141 cttcagacag ctgcttagag cggtttcaga tggacctatt cactgtcact ttgtcccagg
     1201 tcaccttttc cttgggcagc atggttttgg aggtgaccag gcctctctcc aagtggctga
     1261 agcaccctgg ggccctggag aagcagatgt ccagggtagc tggagagtgc tggccgcggc
     1321 cccccacacc gcctgccacc aatgtgctcc ttatgctcta ctccaacctc tcgcaggagc
     1381 agaggcagct gggtgggtcc accttgctgt gggaagccga gagctcctgg cgggcccagg
     1441 agggacagct gtcctgggag tggggcaaga ggcaccgtcg acatgagcag aaactcatct
     1501 cagaagagga tctgcatcac ttgccagaca gaagtcaact gtgtcggaag gtcaagttcc
     1561 aggtggactt caacctgatc ggatggggct cctggatcat ctaccccaag cagtacaacg
     1621 cctatcgctg tgagggcgag tgtcctaatc ctgttgggga ggagtttcat ccgaccaacc
     1681 atgcatacat ccaggtggcc ctgccctgct gtccaaggtc atatgggacc aaaatgtttt
     1741 cattttactc catgaagtct ggaatgagaa tttcttggac ttgcaatata tcctccatgc
     1801 caagcctcag agtctgctga cgtacgcggc cgctcgagca gaaactcatc tcagaagagg
     1861 atctggcagc aaatgatatc ctggattaca aggatgacga cgataaggtt taaacggccg
     1921 gccgcggtca tagctgtttc ctgaacagat cccgggtggc atccctgtga cccctcccca
     1981 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     2041 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     2101 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     2161 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     2221 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     2281 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     2341 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     2401 gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag
     2461 gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc
     2521 ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt
     2581 acgcggccag cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2641 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2701 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2761 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2821 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2881 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2941 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3001 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3061 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3121 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3181 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3241 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3301 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3361 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3421 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat
     3481 ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     3541 gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt
     3601 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     3661 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     3721 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc
     3781 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     3841 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
     3901 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     3961 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     4021 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcgatc cgccacaccc
     4081 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     4141 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg
     4201 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     4261 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     4321 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     4381 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     4441 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     4501 gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg
     4561 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     4621 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     4681 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     4741 tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     4801 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     4861 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     4921 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     4981 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5041 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc
     5101 gcctcaggac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5161 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5221 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5281 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5341 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5401 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5461 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5521 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5581 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5641 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttccatt
     5701 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     5761 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     5821 cccagtcacg acgttgtaaa acgacggcca gtgccaagct gatctataca ttgaatcaat
     5881 attggcaatt agccatatta gtcattggtt atatagcata aatcaata
//
