LOCUS       sequence-222489-        3995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro transcription of human NODAL open reading frame 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
FEATURES             Location/Qualifiers
     source          1..3995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..22
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    139..160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(175..180,181..198,199..205)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    213..229
                     /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     218..240
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     237..253
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     237..253
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     272..291
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        272..290
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           303..345
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           1429..1554
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(1606..1624)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1607..1624)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1632..1649)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1632..1648)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1641..1663)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1857..1876)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1976..1998
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2036..2054)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2122..2226
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2227..2295,2296..3087)
                     /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(2445..2464)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3258..3846
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3747..3766
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
       61 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      121 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      181 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      241 aaacagctat gaccatgatt acgccaagct ctaatacgac tcactatagg gagacaagct
      301 tgcttgttct ttttgcagaa gctcagaata aacgctcaac tttggcagat ctatgcacgc
      361 ccactgcctg cccttccttc tgcacgcctg gtgggcccta ctccaggcgg gtgctgcgac
      421 ggtggccact gcgctcctgc gtacgcgggg gcagccctcg tcgccatccc ctctggcgta
      481 catgctgagc ctctaccgcg acccgctgcc gagggcagac atcatccgca gcctacaggc
      541 agaagatgtg gcagtggatg ggcagaactg gacgtttgct tttgacttct ccttcctgag
      601 ccaacaagag gatctggcat gggctgagct ccggctgcag ctgtccagcc ctgtggacct
      661 ccccactgag ggctcacttg ccattgagat tttccaccag ccaaagcccg acacagagca
      721 ggcttcagac agctgcttag agcggtttca gatggaccta ttcactgtca ctttgtccca
      781 ggtcaccttt tccttgggca gcatggtttt ggaggtgacc aggcctctct ccaagtggct
      841 gaagcgccct ggggccctgg agaagcagat gtccagggta gctggagagt gctggccgcg
      901 gccccccaca ccgcctgcca ccaatgtgct ccttatgctc tactccaacc tctcgcagga
      961 gcagaggcag ctgggtgggt ccaccttgct gtgggaagcc gagagctcct ggcgggccca
     1021 ggagggacag ctgtcctggg agtggggcaa gaggcaccgt cgacatcact tgccagacag
     1081 aagtcaactg tgtcggaagg tcaagttcca ggtggacttc aacctgatcg gatggggctc
     1141 ctggatcatc taccccaagc agtacaacgc ctatcgctgt gagggcgagt gtcctaatcc
     1201 tgttggggag gagtttcatc cgaccaacca tgcatacatc cagagtctgc tgaaacgtta
     1261 ccagccccac cgagtccctt ccacttgttg tgccccagtg aagaccaagc cgctgagcat
     1321 gctgtatgtg gataatggca gagtgctcct agatcaccat aaagacatga tcgtggaaga
     1381 atgtgggtgc ctctgaacta gtgactgact aggatctggt taccactaaa ccagcctcaa
     1441 gaacacccga atggagtctc taagctacat aataccaact tacactttac aaaatgttgt
     1501 cccccaaaat gtagccattc gtatctgctc ctaataaaaa gaaagtttct tcaccccccc
     1561 tgcaggtcga ctctagagga tccccgggcg agctcgaatt cgtattctat agtgtcacct
     1621 aaatccaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     1681 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     1741 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     1801 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     1861 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     1921 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     1981 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2041 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2101 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2161 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2221 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2281 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2341 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     2401 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     2461 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2521 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2581 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2641 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2701 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2761 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2821 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2881 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2941 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3001 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3061 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3121 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3181 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3241 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3301 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3361 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3421 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3481 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3541 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3601 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3661 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3721 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3781 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3841 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3901 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3961 tgagctgata ccgctcgccg cagccgaacg accga
//
