LOCUS       sequence-317062-        5715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription construct for production of cytosolic 
            acvr2b-CIB1N mRNA .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5715)
  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 5715)
  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-60
FEATURES             Location/Qualifiers
     source          1..5715
                     /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             2161..2670
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     polyA_signal    2745..2879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2769..2788)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2823..2842
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2992..3012)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2992..3010)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3031..3047)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3031..3047)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3044..3066)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3055..3071
                     /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(3079..3085,3086..3103,3104..3109))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3124..3145
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3262..3279)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3433..4021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3513..3532)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4192..4983,4984..5052))
                     /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     4815..4834
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5053..5157)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5183..5638)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5320..5341)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5532..5551
                     /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 ccatgaaacc tccgtacgga
     1081 catgtggacg tcaatgagga tccaggccca tctcctccat ctcctctggt gggtctgaag
     1141 cctctgcagc tgctggaggt taaagctcgc ggacgcttcg gctgcgtctg gaaggctcag
     1201 atgatcaatg aatatgtagc tgtcaagatt ttccccattc aggataagct gtcgtggcag
     1261 aacgagcggg agatgttttc cactccggga atgaaacatg ataacctgct gcgcttcatc
     1321 gctgctgaga aacgcggatc taacctggag atggagttct ggctcatcac tgagtttcat
     1381 gagcggggct ctctgacgga ctatctgaag gggaacgcag tgagctgggc tgatctgtgt
     1441 gtgatagcgg agagcatggc ctgtggtctg gcgtatctgc atgaagacgt gccgcgctcc
     1501 aaaggagaag gccccaaacc agccatcgca cacagagact tcaagagcaa gaatgtgatg
     1561 ctgaagatgg acctcaccgc cgtcattggg gattttgggc tggcggtgcg gtttgagccg
     1621 gggaaaccgc cgggagacac acatggccag gtgggcacga ggaggtacat ggccccggag
     1681 gttctggaag gagccataaa cttccagcgg gactcctttc tgcggataga catgtacgcc
     1741 atgggcctgg tgctgtggga gctggtgtca cgctgcaaag ctgctgatgg tcctgtggac
     1801 gagtacatgc tgccgtttga ggaggagatc ggtcagcacc cgtcgctgga ggatctgcag
     1861 gatgctgtgg tccataagaa gctgcggccg gcgtttaagg actgctggct caagcattca
     1921 ggtctgtgtc agatgtgcga gaccatggag gagtgctggg atcatgacgc agaggctcgt
     1981 ctgtcggccg gctgtgtgca ggagcgcatc tctcagatcc gccgcgtcag cagctccacc
     2041 tcagactgcc tgttctccat ggtgacctcg ctcaccaacg tggacctgcc gcccaaagag
     2101 tccagcatcg gcggcggcgg cagtggcggc ggcggctctg gcggcggcgg ctcttctaga
     2161 atgaatggag ctataggagg tgaccttttg ctcaattttc ctgacatgtc ggtcctagag
     2221 cgccaaaggg ctcacctcaa gtacctcaat cccacctttg attctcctct cgccggcttc
     2281 tttgccgatt cttcaatgat taccggcggc gagatggaca gctatctttc gactgccggt
     2341 ttgaatcttc cgatgatgta cggtgagacg acggtggaag gtgattcaag actctcaatt
     2401 tcgccggaaa cgacgcttgg gactggaaat ttcaaggcag cgaagtttga tacagagact
     2461 aaggattgta atgaggcggc gaagaagatg acgatgaaca gagatgacct agtagaagaa
     2521 ggagaagaag agaagtcgaa aataacagag caaaacaatg ggagcacaaa aagcatcaag
     2581 aagatgaaac acaaagccaa gaaagaagag aacaatttct ctaatgattc atctaaagtg
     2641 acgaaggaat tggagaaaac ggattatatt catgtaccgg tcgccaccat gtaagaattc
     2701 aaggcctctc gagcctcaag aactatagtg agtcgtatta cgtagatcca gacatgataa
     2761 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2821 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2881 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     2941 aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg
     3001 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3061 tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc
     3121 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3181 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3241 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3301 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3361 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3421 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3481 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3541 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3601 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3661 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3721 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3781 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3841 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3901 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3961 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4021 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4081 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4141 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4201 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4261 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4321 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4381 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4441 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4501 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4561 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4621 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4681 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4741 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4801 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4861 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4921 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4981 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5041 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5101 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5161 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     5221 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     5281 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     5341 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     5401 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     5461 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     5521 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     5581 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     5641 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5701 ttacgccagt cgacc
//
