LOCUS       Exported                5991 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  For mammalian expression of a C-terminal truncated human NODAL open 
            reading frame with an internal MYC tag.
ACCESSION   .
VERSION     .
KEYWORDS    NODAL-matMYC-ex2trunc
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   bioRxiv 276170
REFERENCE   2  (bases 1 to 5991)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5991
                     /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             1743..1772
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2157..2186
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2205..2228
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2248..2264)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2248..2264)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2275..2897
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2474..2493)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(3046..3634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3126..3145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3963..4010
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4035..4054
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4242..5036)
                     /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(4353..4372)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4963..4982
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5071..5428)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5085..5220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5139..5158)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5385..5405
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5430..5534)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5561..25
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5648..5667)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5858..5879
                     /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 atgagcagaa actcatctca gaagaggatc tgcatcactt gccagacaga agtcaactgt
     1801 gtcggaaggt caagttccag gtggacttca acctgatcgg atggggctcc tggatcatct
     1861 accccaagca gtacaacgcc tatcgctgtg agggcgagtg tcctaatcct gttggggagg
     1921 agtttcatcc gaccaaccat gcatacatcc agtgactgct gaaacgttac cagcatgcat
     1981 acatccagtg actgctgaaa cgttaccagc cccaccgagt cccttccact tgttgtgccc
     2041 cagtgaagac caagccgctg agcatgctgt atgtggataa tggcagagtg ctcctagatc
     2101 accataaaga catgatcgtg gaagaatgtg ggtgcctctg acgtacgcgg ccgctcgagc
     2161 agaaactcat ctcagaagag gatctggcag caaatgatat cctggattac aaggatgacg
     2221 acgataaggt ttaaacggcc ggccgcggtc atagctgttt cctgaacaga tcccgggtgg
     2281 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     2341 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     2401 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     2461 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     2521 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     2581 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     2641 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     2701 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta
     2761 aaataactat accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa
     2821 ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag
     2881 tagatgcctg ttgaattggg tacgcggcca gcggcgagcg gtatcagctc actcaaaggc
     2941 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3001 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3061 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3121 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3181 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3241 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3301 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3361 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3421 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3481 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3541 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3601 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3661 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3721 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3781 atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg
     3841 gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc
     3901 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt
     3961 atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg
     4021 ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa
     4081 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     4141 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     4201 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     4261 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     4321 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     4381 tgatagcgat ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     4441 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     4501 ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     4561 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     4621 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     4681 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     4741 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     4801 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt
     4861 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     4921 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     4981 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     5041 aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc
     5101 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     5161 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     5221 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     5281 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     5341 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     5401 attccacagc tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt
     5461 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5521 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     5581 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5641 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5701 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5761 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5821 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5881 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5941 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt t
//
