LOCUS       sequence-222542-        3995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro transcription of human NODAL open reading frame (with 
            two mutated N-glycosylation sites) RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3995)
  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 3995)
  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-T7TS-N72A-N199A
FEATURES             Location/Qualifiers
     source          1..3995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    63..84
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(99..104,105..122,123..129)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    137..153
                     /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     142..164
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     161..177
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     161..177
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     196..215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        196..214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           227..269
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           1353..1478
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(1530..1548)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1531..1548)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1556..1573)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1556..1572)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1565..1587)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1781..1800)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1900..1922
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1960..1978)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2046..2150
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2151..2219,2220..3011)
                     /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(2369..2388)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3182..3770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3671..3690
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3924..3941
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
       61 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      121 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      181 gattacgcca agctctaata cgactcacta tagggagaca agcttgcttg ttctttttgc
      241 agaagctcag aataaacgct caactttggc agatctatgc acgcccactg cctgcccttc
      301 cttctgcacg cctggtgggc cctactccag gcgggtgctg cgacggtggc cactgcgctc
      361 ctgcgtacgc gggggcagcc ctcgtcgcca tcccctctgg cgtacatgct gagcctctac
      421 cgcgacccgc tgccgagggc agacatcatc cgcagcctac aggcagaaga tgtggcagtg
      481 gatgggcagg cctggacgtt tgcttttgac ttctccttcc tgagccaaca agaggatctg
      541 gcatgggctg agctccggct gcagctgtcc agccctgtgg acctccccac tgagggctca
      601 cttgccattg agattttcca ccagccaaag cccgacacag agcaggcttc agacagctgc
      661 ttagagcggt ttcagatgga cctattcact gtcactttgt cccaggtcac cttttccttg
      721 ggcagcatgg ttttggaggt gaccaggcct ctctccaagt ggctgaagcg ccctggggcc
      781 ctggagaagc agatgtccag ggtagctgga gagtgctggc cgcggccccc cacaccgcct
      841 gccaccaatg tgctccttat gctctactcc gccctctcgc aggagcagag gcagctgggt
      901 gggtccacct tgctgtggga agccgagagc tcctggcggg cccaggaggg acagctgtcc
      961 tgggagtggg gcaagaggca ccgtcgacat cacttgccag acagaagtca actgtgtcgg
     1021 aaggtcaagt tccaggtgga cttcaacctg atcggatggg gctcctggat catctacccc
     1081 aagcagtaca acgcctatcg ctgtgagggc gagtgtccta atcctgttgg ggaggagttt
     1141 catccgacca accatgcata catccagagt ctgctgaaac gttaccagcc ccaccgagtc
     1201 ccttccactt gttgtgcccc agtgaagacc aagccgctga gcatgctgta tgtggataat
     1261 ggcagagtgc tcctagatca ccataaagac atgatcgtgg aagaatgtgg gtgcctctga
     1321 actagtgact gactaggatc tggttaccac taaaccagcc tcaagaacac ccgaatggag
     1381 tctctaagct acataatacc aacttacact ttacaaaatg ttgtccccca aaatgtagcc
     1441 attcgtatct gctcctaata aaaagaaagt ttcttcaccc cccctgcagg tcgactctag
     1501 aggatccccg ggcgagctcg aattcgtatt ctatagtgtc acctaaatcc aattcactgg
     1561 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     1621 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     1681 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
     1741 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     1801 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     1861 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1921 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     1981 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     2041 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2101 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2161 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2221 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2281 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2341 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2401 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2461 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2521 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2581 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2641 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2701 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2761 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2821 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2881 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2941 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3001 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3061 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3121 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3181 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3241 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3301 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3361 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3421 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3481 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3541 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3601 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3661 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3721 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3781 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3841 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3901 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     3961 tacgcaaacc gcctctcccc gcgcgttggc cgatt
//
