LOCUS       Exported                5895 bp ds-DNA     circular SYN 13-MAY-2021
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    NODALvar-MYC-DYK-C302A
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   bioRxiv 276170
REFERENCE   2  (bases 1 to 5895)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5895
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2061..2090
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2109..2132
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2152..2168)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2152..2168)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2179..2801
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2378..2397)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2950..3538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3030..3049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3867..3914
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3939..3958
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4146..4940)
                     /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(4257..4276)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4867..4886
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4975..5332)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4989..5124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5043..5062)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5289..5309
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5334..5438)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5465..25
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5552..5571)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5762..5783
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcatcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat gcacgcccac tgcctgccct tccttctgca cgcctggtgg gccctactcc
     1081 aggcgggtgc tgcgacggtg gccactgcgc tcctgcgtac gcgggggcag ccctcgtcgc
     1141 catcccctct ggcgtacatg ctgagcctct accgcgaccc gctgccgagg gcagacatca
     1201 tccgcagcct acaggcagaa gatgtggcag tggatgggca gaactggacg tttgcttttg
     1261 acttctcctt cctgagccaa caagaggatc tggcatgggc tgagctccgg ctgcagctgt
     1321 ccagccctgt ggacctcccc actgagggct cacttgccat tgagattttc caccagccaa
     1381 agcccgacac agagcaggct tcagacagct gcttagagcg gtttcagatg gacctattca
     1441 ctgtcacttt gtcccaggtc accttttcct tgggcagcat ggttttggag gtgaccaggc
     1501 ctctctccaa gtggctgaag caccctgggg ccctggagaa gcagatgtcc agggtagctg
     1561 gagagtgctg gccgcggccc cccacaccgc ctgccaccaa tgtgctcctt atgctctact
     1621 ccaacctctc gcaggagcag aggcagctgg gtgggtccac cttgctgtgg gaagccgaga
     1681 gctcctggcg ggcccaggag ggacagctgt cctgggagtg gggcaagagg caccgtcgac
     1741 atcacttgcc agacagaagt caactgtgtc ggaaggtcaa gttccaggtg gacttcaacc
     1801 tgatcggatg gggctcctgg atcatctacc ccaagcagta caacgcctat cgctgtgagg
     1861 gcgagtgtcc taatcctgtt ggggaggagt ttcatccgac caaccatgca tacatccagg
     1921 tggccctgcc cgcctgtcca aggtcatatg ggaccaaaat gttttcattt tactccatga
     1981 agtctggaat gagaatttct tggacttgca atatatcctc catgccaagc ctcagagtct
     2041 gcacgcgtac gcggccgctc gagcagaaac tcatctcaga agaggatctg gcagcaaatg
     2101 atatcctgga ttacaaggat gacgacgata aggtttaaac ggccggccgc ggtcatagct
     2161 gtttcctgaa cagatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     2221 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     2281 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     2341 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     2401 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     2461 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     2521 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     2581 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     2641 cccttccctg tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag
     2701 cataggctac ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc
     2761 ctctcatgcg ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcggcg
     2821 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2881 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2941 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3001 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3061 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3121 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3181 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3241 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3301 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3361 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3421 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3481 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3541 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3601 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3661 aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc
     3721 cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa
     3781 ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca
     3841 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg
     3901 tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt
     3961 tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
     4021 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
     4081 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     4141 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     4201 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     4261 tcacgggtag ccaacgctat gtcctgatag cgatccgcca cacccagccg gccacagtcg
     4321 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     4381 gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct
     4441 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     4501 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     4561 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     4621 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     4681 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     4741 agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     4801 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     4861 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     4921 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca
     4981 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     5041 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     5101 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     5161 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     5221 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5281 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc
     5341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5461 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5521 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5581 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5641 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5701 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5761 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5821 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5881 tttaacgcga atttt
//
