LOCUS       sequence-222491-        3979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro transcription of a human NODAL splice variant open 
            reading frame RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3979)
  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 3979)
  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-T7TS
FEATURES             Location/Qualifiers
     source          1..3979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     54..71
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    188..209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(224..229,230..247,248..254)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    262..278
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     267..289
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     286..302
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     286..302
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     321..340
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        321..339
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           352..394
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           1451..1576
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(1639..1657)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1640..1657)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1665..1682)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1665..1681)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1674..1696)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1890..1909)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2009..2031
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2069..2087)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2155..2259
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2260..2328,2329..3120)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2478..2497)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3291..3879
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3780..3799
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
       61 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
      121 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
      181 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
      241 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
      301 accatgatta cgccaagctc taatacgact cactataggg agacaagctt gcttgttctt
      361 tttgcagaag ctcagaataa acgctcaact ttggcagatc tatgcacgcc cactgcctgc
      421 ccttccttct gcacgcctgg tgggccctac tccaggcggg tgctgcgacg gtggccactg
      481 cgctcctgcg tacgcggggg cagccctcgt cgccatcccc tctggcgtac atgctgagcc
      541 tctaccgcga cccgctgccg agggcagaca tcatccgcag cctacaggca gaagatgtgg
      601 cagtggatgg gcagaactgg acgtttgctt ttgacttctc cttcctgagc caacaagagg
      661 atctggcatg ggctgagctc cggctgcagc tgtccagccc tgtggacctc cccactgagg
      721 gctcacttgc cattgagatt ttccaccagc caaagcccga cacagagcag gcttcagaca
      781 gctgcttaga gcggtttcag atggacctat tcactgtcac tttgtcccag gtcacctttt
      841 ccttgggcag catggttttg gaggtgacca ggcctctctc caagtggctg aagcaccctg
      901 gggccctgga gaagcagatg tccagggtag ctggagagtg ctggccgcgg ccccccacac
      961 cgcctgccac caatgtgctc cttatgctct actccaacct ctcgcaggag cagaggcagc
     1021 tgggtgggtc caccttgctg tgggaagccg agagctcctg gcgggcccag gagggacagc
     1081 tgtcctggga gtggggcaag aggcaccgtc gacatcactt gccagacaga agtcaactgt
     1141 gtcggaaggt caagttccag gtggacttca acctgatcgg atggggctcc tggatcatct
     1201 accccaagca gtacaacgcc tatcgctgtg agggcgagtg tcctaatcct gttggggagg
     1261 agtttcatcc gaccaaccat gcatacatcc aggtggccct gccctgctgt ccaaggtcat
     1321 atgggaccaa aatgttttca ttttactcca tgaagtctgg aatgagaatt tcttggactt
     1381 gcaatatatc ctccatgcca agcctcagag tctgctgaac tagtgactga ctaggatctg
     1441 gttaccacta aaccagcctc aagaacaccc gaatggagtc tctaagctac ataataccaa
     1501 cttacacttt acaaaatgtt gtcccccaaa atgtagccat tcgtatctgc tcctaataaa
     1561 aagaaagttt cttcacattc taaaaacccc ccctgcaggt cgactctaga ggatccccgg
     1621 gcgagctcga attcgtattc tatagtgtca cctaaatcca attcactggc cgtcgtttta
     1681 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     1741 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     1801 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     1861 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     1921 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     1981 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2041 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2101 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2161 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2221 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2281 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2341 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2401 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2461 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2521 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2581 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2641 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2701 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2761 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2821 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2881 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2941 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3001 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3061 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3121 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3181 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3241 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3301 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3361 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3421 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3481 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3541 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3601 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3661 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3721 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3781 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3841 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3901 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3961 gattctgtgg ataaccgta
//
