LOCUS       sequence-317070-        6789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription construct for production of membrane 
            acvr1b-Cry2 mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6789)
  AUTHORS   McNamara HM, Jia BZ, Guyer A, Parot VJ, Dobbs C, Schier AF, Cohen 
            AE, Lord ND
  TITLE     Optogenetic control of Nodal signaling patterns.
  JOURNAL   bioRxiv [Preprint]. 2024 Apr 12:2024.04.11.588875. doi: 
            10.1101/2024.04.11.588875.
  PUBMED    38645239
REFERENCE   2  (bases 1 to 6789)
  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: "bioRxiv 
            [Preprint]. 2024 Apr 12:2024.04.11.588875. doi: 
            10.1101/2024.04.11.588875."
COMMENT     Sequence Label: pNDL-54
FEATURES             Location/Qualifiers
     source          1..6789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1063..1104
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             2245..3738
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     polyA_signal    3819..3953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3843..3862)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3897..3916
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4105..4121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4105..4121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4118..4140)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4129..4145
                     /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)."
     promoter        complement(join(4153..4159,4160..4177,4178..4183))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4198..4219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4336..4353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4507..5095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4587..4606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5266..6057,6058..6126))
                     /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     5889..5908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6127..6231)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6257..6712)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6394..6415)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6606..6625
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat ccatggggag tagcaagagc
     1081 aagcctaagg accccagcca gcgcggtgga ggaggttctg gaggcggtgg aagtggtggc
     1141 ggaggtagcc accggcagcg actggatgtt gaggatccat cctgtgatca cctgtacttg
     1201 gccaaagaca agaccttaca ggatctcatc ttcgatctgt ccacctctgg ttcagggtct
     1261 gggctgccct tgttcgttca gaggactgtg gccaggacaa ttgtactgca ggagatcata
     1321 ggaaaaggtc gctttgggga agtgtggagg gggagatgga gaggaggtga tgtggctgtg
     1381 aagatcttct catccaggga ggaacgttcc tggttccgtg aagctgagat ttaccaaacc
     1441 atcatgctcc gccatgaaaa catcttgggc ttcattgctg ctgataataa agacaatggc
     1501 acatggacac agctgtggct agtgtcagac taccatgagc atggctcatt atttgactat
     1561 ctcaaccact actccgtcac aatcgagggg atgatcaagt tatcgctttc agccgccagc
     1621 ggtctggcac atctgcacat ggagatcctg ggcacacagg ggaaaccagg catcgcacat
     1681 cgtgacctca aatccaaaaa catcttggtg aaaaagaatg gtacttgcgc catagcagat
     1741 ctagggctcg ctgtacggca cgagtccatc actgatacta tagacattgc acccaatcag
     1801 agggtgggca ctaaaagata tatggcccca gaggtactag atgaaacgat caacatgaag
     1861 cattttgatt ctttcaagtg tgctgatatc tatgctttgg gactggtata ttgggagatt
     1921 gcacgacggt gtaatgctgg aggtatccat gaagattatc agctacccta ctatgacctg
     1981 gtgccgtccg acccctccat agaagagatg aggaaggtgg tgtgtgacca gaggcttcgg
     2041 cctaatgtgc ccaactggtg gcagagctat gaggcgctga gagtgatggg aaagatcatg
     2101 cgggagtgtt ggtatgccaa cggagcggcg cggcttacag ctctgcggat taagaagact
     2161 ctttcgcaac tcagcgtcca agaagacatt aagggtggag gaggttctgg aggcggtgga
     2221 agtggtggcg gaggtagctc tagaatgaag atggacaaaa agactatagt ttggtttaga
     2281 agagacctaa ggattgagga taatcctgca ttagcagcag ctgctcacga aggatctgtt
     2341 tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg aagagcttca
     2401 agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc tcttggatct
     2461 gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg tatccgcgtt
     2521 accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt agttcgggac
     2581 cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta caatggagat
     2641 ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac gagtttcaat
     2701 tcttactgga agaaatgctt agatatgtcg attgaatccg ttatgcttcc tcctccttgg
     2761 cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat tgaagaacta
     2821 gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc ttggtctcca
     2881 ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt gatagattat
     2941 gcaaagaaca gcaagaaagt tgttgggaat tctacttcac tactttctcc gtatctccat
     3001 ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca aattatatgg
     3061 gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag gggaatcggt
     3121 ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga gcaatcgttg
     3181 ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa ggcctggaga
     3241 caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg ggctaccgga
     3301 tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt tcttctcctt
     3361 ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga tttggaatgt
     3421 gacatccttg gctggcagta tatctctggg agtatccccg atggccacga gcttgatcgc
     3481 ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta cataaggcaa
     3541 tggcttcccg agcttgcgag attgccaact gaatggatcc atcatccatg ggacgctcct
     3601 ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa acccattgta
     3661 gacatcgaca cagctcgtga gctactagct aaagctattt caagaacccg tgaagcacag
     3721 atcatgatcg gagcagcagc ccgggatcca ccggtcgcca ccatgtaaga attcaaggcc
     3781 tctcgagcct caagaactat agtgagtcgt attacgtaga tccagacatg ataagataca
     3841 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3901 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3961 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     4021 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtc
     4081 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4141 cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg
     4201 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4261 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4321 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4381 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4441 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4501 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4561 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4621 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4681 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4741 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4801 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4861 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4921 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4981 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5041 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5101 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5161 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5221 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5281 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5341 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5401 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5461 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5521 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5581 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5641 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5701 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5761 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5821 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5881 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5941 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6001 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6061 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6121 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6181 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6241 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     6301 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     6361 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     6421 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     6481 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     6541 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     6601 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     6661 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     6721 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     6781 cagtcgacc
//
