LOCUS       sequence-496437-        6348 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Expresses recombinant fluorescent CYRI-B (Fam49B) protein in 
            mammalian cells, specifically human Cyri-B with internal mCherry 
            after proline 17, preserving the proposed N-terminal myristoylation 
            site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6348)
  AUTHORS   Le AH, Yelland T, Paul NR, Fort L, Nikolaou S, Ismail S, Machesky LM
  TITLE     CYRI-A limits invasive migration through macropinosome formation and
            integrin uptake regulation.
  JOURNAL   J Cell Biol. 2021 Sep 6;220(9). pii: 212437. doi: 
            10.1083/jcb.202012114. Epub 2021 Jun 24.
  PUBMED    34165494
REFERENCE   2  (bases 1 to 6348)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 7, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol."; date: "2021-09-6"; volume: "220(9). pii: 212437. doi"; 
            pages: " 10.1083/jcb.202012114. Epub 2021 Jun 24"
COMMENT     Sequence Label: p17-mCherry-CYRI-B
FEATURES             Location/Qualifiers
     source          1..6348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        1020..1399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1400..1603
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1554..1574
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1600..1624
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1834..1852
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1834..1851
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1865..1884
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1988..2692
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2130..2148)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2398..2417
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      3654..3663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3667..3801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3704..3723)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3758..3777
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3870..4227
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3870..4061
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3893..3913)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4078..4213
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4247..4264)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4247..4263)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4256..4278)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4476..4931
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4563..4582)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4773..4794
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4948..4967)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5067..5089
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5127..5145)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5213..5317
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5318..5386,5387..6178)
                     /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(5536..5555)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgctacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga catgattacg aattaattcg agctcgcccg
     1021 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     1081 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     1141 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     1201 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     1261 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     1321 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     1381 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     1441 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     1501 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     1561 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     1621 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
     1681 cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca agagtgacgt
     1741 aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac gcggctacaa
     1801 ttaatacata accttatgta tcatacacat acgatttagg tgacactata gaataacatc
     1861 cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct cggttctatc
     1921 gattgaattc atggggaatc ttcttaaagt tttgacatgc acagaccttg agcaggggcc
     1981 actcgaggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     2041 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     2101 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     2161 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     2221 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     2281 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     2341 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     2401 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     2461 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     2521 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     2581 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     2641 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca aggctagcaa
     2701 ttttttcctt gattttgaaa atgcccagcc tacagagtct gagaaggaaa tttataatca
     2761 ggtgaatgta gtattaaaag atgcagaagg catcttggag gacttgcagt catacagagg
     2821 agctggccac gaaatacgag aggcaatcca gcatccagca gatgagaagt tgcaagagaa
     2881 ggcatggggt gcagttgttc cactagtagg caaattaaag aaattttacg aattttctca
     2941 gaggttagaa gcagcattaa gaggtcttct gggagcctta acaagtaccc catattctcc
     3001 cacccagcat ctagagcgag agcaggctct tgctaaacag tttgcagaaa ttcttcattt
     3061 cacactccgg tttgatgaac tcaagatgac aaatcctgcc atacagaatg atttcagcta
     3121 ttatagaaga acattgagtc gtatgaggat taacaatgta ccggcagaag gagaaaatga
     3181 agtaaataat gaattggcaa atcgaatgtc tttgttttat gctgaggcaa ctccaatgct
     3241 gaaaaccttg agtgatgcca caacaaaatt tgtatcagag aataaaaatt taccaataga
     3301 aaataccaca gattgtttaa gcacaatggc tagtgtatgc agagtcatgc tggaaacacc
     3361 ggaatacaga agcagattta caaatgaaga gacagtgtca ttctgcttga gggtaatggt
     3421 gggtgtcata atactctatg accacgtaca tccagtggga gcatttgcta aaacttccaa
     3481 aattgatatg aaaggttgta tcaaagttct taaggaccaa cctcctaata gtgtggaagg
     3541 tcttctaaat gctctcaggt acacaacaaa acatttgaat gatgagacta cctccaagca
     3601 aattaaatcc atgctgcaat aaggatcctc tagagtcgac ctgcagaagc ttggccgcca
     3661 tggcccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3721 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3781 gtatcttatc atgtctggat cgggaattaa ttcggcgcag caccatggcc tgaaataacc
     3841 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     3901 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3961 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4021 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4081 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4141 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4201 ttttttggag gcctaggctt ttgcaaaaag ctgttaacag cttggcactg gccgtcgttt
     4261 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     4321 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4381 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     4441 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     4501 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4561 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4621 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4681 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4741 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4801 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     4861 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4921 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4981 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5041 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5101 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5161 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5221 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5281 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5341 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5401 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5461 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5521 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5581 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5641 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5701 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5761 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5821 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5881 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5941 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6001 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6061 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6121 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6181 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6241 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6301 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
