LOCUS       sequence-222540-        3993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro transcription of human NODAL open reading frame (with a
            mutated Cysteine residue) RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3993)
  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 3993)
  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-C312S
FEATURES             Location/Qualifiers
     source          1..3993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..34
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    151..172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(187..192,193..210,211..217)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    225..241
                     /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     230..252
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     249..265
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     249..265
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     284..303
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        284..302
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           315..357
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           1441..1566
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(1616..1634)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1617..1634)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1642..1659)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1642..1658)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1651..1673)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1867..1886)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1986..2008
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2046..2064)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2132..2236
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2237..2305,2306..3097)
                     /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(2455..2474)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3268..3856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3757..3776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
       61 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
      121 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
      181 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
      241 atttcacaca ggaaacagct atgaccatga ttacgccaag ctctaatacg actcactata
      301 gggagacaag cttgcttgtt ctttttgcag aagctcagaa taaacgctca actttggcag
      361 atctatgcac gcccactgcc tgcccttcct tctgcacgcc tggtgggccc tactccaggc
      421 gggtgctgcg acggtggcca ctgcgctcct gcgtacgcgg gggcagccct cgtcgccatc
      481 ccctctggcg tacatgctga gcctctaccg cgacccgctg ccgagggcag acatcatccg
      541 cagcctacag gcagaagatg tggcagtgga tgggcagaac tggacgtttg cttttgactt
      601 ctccttcctg agccaacaag aggatctggc atgggctgag ctccggctgc agctgtccag
      661 ccctgtggac ctccccactg agggctcact tgccattgag attttccacc agccaaagcc
      721 cgacacagag caggcttcag acagctgctt agagcggttt cagatggacc tattcactgt
      781 cactttgtcc caggtcacct tttccttggg cagcatggtt ttggaggtga ccaggcctct
      841 ctccaagtgg ctgaagcgcc ctggggccct ggagaagcag atgtccaggg tagctggaga
      901 gtgctggccg cggcccccca caccgcctgc caccaatgtg ctccttatgc tctactccaa
      961 cctctcgcag gagcagaggc agctgggtgg gtccaccttg ctgtgggaag ccgagagctc
     1021 ctggcgggcc caggagggac agctgtcctg ggagtggggc aagaggcacc gtcgacatca
     1081 cttgccagac agaagtcaac tgtgtcggaa ggtcaagttc caggtggact tcaacctgat
     1141 cggatggggc tcctggatca tctaccccaa gcagtacaac gcctatcgct gtgagggcga
     1201 gtgtcctaat cctgttgggg aggagtttca tccgaccaac catgcataca tccagagtct
     1261 gctgaaacgt taccagcccc accgagtccc ttccacttct tgtgccccag tgaagaccaa
     1321 gccgctgagc atgctgtatg tggataatgg cagagtgctc ctagatcacc ataaagacat
     1381 gatcgtggaa gaatgtgggt gcctctgaac tagtgactga ctaggatctg gttaccacta
     1441 aaccagcctc aagaacaccc gaatggagtc tctaagctac ataataccaa cttacacttt
     1501 acaaaatgtt gtcccccaaa atgtagccat tcgtatctgc tcctaataaa aagaaagttt
     1561 cttcacattc taaaaaaaga ctctagagga tccccgggcg agctcgaatt cgtattctat
     1621 agtgtcacct aaatccaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1681 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1741 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     1801 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     1861 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     1921 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     1981 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     2041 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2101 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2161 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2221 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2281 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2341 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2401 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2461 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2521 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2581 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2641 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2701 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2761 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2821 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2881 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2941 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3001 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3061 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3121 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3181 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3241 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3301 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3361 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3421 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3481 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3541 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3601 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3661 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3721 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3781 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3841 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3901 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3961 cgcctttgag tgagctgata ccgctcgccg cag
//
