LOCUS       sequence-367398-        5382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRNA synthesis of rapalog-induced cell adhesion molecule.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5382)
  AUTHORS   Usami C, Inomata H
  TITLE     Rapalog-induced cell adhesion molecule inhibits mesoderm migration 
            in Xenopus embryos by increasing frequency of adhesion to the 
            ectoderm.
  JOURNAL   Genes Cells. 2022 Jun;27(6):436-450. doi: 10.1111/gtc.12937. Epub 
            2022 May 4.
  PUBMED    35437867
REFERENCE   2  (bases 1 to 5382)
  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"; doi: "10.1111/gtc.12937";
            journalName: "Genes Cells"; date: "2022-06"; volume: "27"; issue: 
            "6"; pages: "436-450"
COMMENT     Sequence Label: pCS2-SP-FKBP-mCherry-TM-V5
FEATURES             Location/Qualifiers
     source          1..5382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        861..879
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     861..878
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1001..1321
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1322..2029
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1467..1485)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1735..1754
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2186..2227
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    2260..2394
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2284..2303)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2338..2357
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2507..2527)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2507..2525)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2546..2562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2546..2562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2559..2581)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2570..2586
                     /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(2594..2600,2601..2618,2619..2624))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2639..2660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2777..2794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2948..3536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3028..3047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3707..4498,4499..4567))
                     /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     4330..4349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4568..4672)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4698..5153)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4835..4856)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5047..5066
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5224..5382,1..826)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact tggcacggtg
       61 ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg
      121 ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat
      181 ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata
      241 tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc
      301 atataccata tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg
      361 gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc
      421 gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt
      481 caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc
      541 tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct
      601 attaatagta acttggcaag tacattacta ttggaagtac gccagggtac attggcagta
      661 ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg
      721 acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag
      781 gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaaccgcc attctgcctg
      841 gggacgtcgg agcaagcttg atttaggtga cactatagaa tacaagctac ttgttctttt
      901 tgcaggatcc gccatgaact ccttctccac aagcgccttc ggtccagttg ccttctccct
      961 gggcctgctc ctggtgttgc ctgctgcctt ccctgcccca ggagtgcagg tggaaaccat
     1021 ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgcgtgg tgcactacac
     1081 cgggatgctt gaagatggaa agaaatttga ttcctcccgg gacagaaaca agccctttaa
     1141 gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg cccagatgag
     1201 tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg ccactgggca
     1261 cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc taaaactgga
     1321 aatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     1381 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     1441 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     1501 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     1561 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     1621 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     1681 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     1741 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     1801 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     1861 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     1921 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     1981 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg agctgttttg
     2041 ggcactggtc gtggttgctg gagtcctgtt ttgttatggc ttgctagtga cagtggctct
     2101 ttgtgttatc tggacaaata gtagaaggaa cagaggtggc tctggcggtg gcggatcggg
     2161 tggaggttcg tccggaggcc ccgggggtaa gcctatccct aaccctctcc tcggtctcga
     2221 ttctacgtag tctagaacta tagtgagtcg tattacgtag atccagacat gataagatac
     2281 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2341 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2401 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc
     2461 gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt
     2521 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2581 tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg ggtgcctaat
     2641 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2701 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2761 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2821 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2881 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2941 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3001 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3061 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3121 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3181 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3241 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3301 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3361 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3421 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3481 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3541 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3601 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3661 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3721 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3781 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3841 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3901 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3961 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4021 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4081 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4141 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4201 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4261 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4321 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4381 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4441 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4501 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4561 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4621 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4681 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     4741 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4801 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4861 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4921 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4981 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     5041 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     5101 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     5161 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5221 ccagtcgacc gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat
     5281 ctggacctgt gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat
     5341 ctggacccca gccaattcaa tatggcggac ttggcaccat gc
//
