LOCUS       sequence-209765-        5445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mEGFP-linker sequence for N-terminus tagging of 
            human CTNNB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5445)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5445)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AICSDP-47: CTNNB1-mEGFP
FEATURES             Location/Qualifiers
     source          1..5445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     180..202
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     194..211
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     195..211
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1229..1231,1232..1234,1235..1945)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1274..1295)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1535..1554)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1882..1903
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2492..2527
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     primer_bind     complement(3040..3056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3040..3056)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3053..3075)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3064..3080
                     /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(3088..3094,3095..3112,3113..3118))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3133..3154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3271..3288)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3442..4030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3522..3541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4201..4992,4993..5061))
                     /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     4824..4843
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5062..5166)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5234..5252
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5290..5312)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5412..5431
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
       61 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      121 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      181 ccagtcacga cgttgtaaaa cgacggccag tgaattgacg cgtattggga tctcaaaaaa
      241 aaacaaaaca tagttcacac ttaaatattt tattccatat ctttacatac ccaatatgtt
      301 aatttatagt tcaagatgaa cttgtttggg acagattttg taataaagga aatcgtgtta
      361 ttagaaatat ctagaggcca tgagccctta aactgttcta atttgcaagt agttccctgt
      421 gtgatgcagt ttttttcaat attgcacaat aaaggcaaaa tacggacaaa ttagatgata
      481 agatttatat aaatttttaa aatattgatc aaaatatgta tccatattgg taatatttgt
      541 atttataata aatcattgct gtaaatttga acttagaaaa attttactaa taaaggtgct
      601 tttgtgttgc aaactttcat ttgaaaagta atttttcttt gtaccaaaaa atctaaaatt
      661 cgctattcta gtcaccaaaa tttgctttat gaaaaataat ttttgatggc actatatcag
      721 aaaacaactt gttaaagaaa atgtggagtt tttaaaatcc cactgtacct ctgttatcca
      781 aaggggatct gtgaattttt ctgtgaaagg ttaaaaaagg agagaccttt aggaattcag
      841 agagcagctg atttttgaat agtgttttcc cctccctggc ttttattatt acaactctgt
      901 gctttttcat caccatcctg aatatctata attaatattt atactattaa taaaaagaca
      961 tttttggtaa ggaggagttt tcactgaagt tcagcagtga tggagctgtg gttgaggtgt
     1021 ctggaggaga ccatgaggtc tgcgtttcac taacctggta aaagaggata tgggtttttt
     1081 ttgtgggtgt aatagtgaca tttaacaggt atcccagtga cttaggagta ttaatcaagc
     1141 taaatttaaa tcctaatgac ttttgattaa ctttttttag ggtatttgaa gtataccata
     1201 caactgtttt gaaaacccgg tgtggacaat ggtgagcaag ggcgaggagc tgttcaccgg
     1261 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1321 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1381 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     1441 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1501 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1561 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1621 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1681 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     1741 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1801 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga
     1861 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1921 tctcggcatg gacgagctgt acaagagcgg cctgagaagc agagcccagg ccagcaacag
     1981 cgccgtggac ggaaccgcag caaccgctac tcaaggtttg tgtcattaaa tctttagtta
     2041 ctgaattggg gctctgcttc gttgccatta agccagtctg gctgagatcc ccctgctttc
     2101 ctctctccct gcttacttgt caggctacct tttgctccat tttctgctca ctcctcctaa
     2161 tggcttggtg aaatagcaaa caagccacca gcaggaatct agtctggatg actgcttctg
     2221 gagcctggat gcagtaccat tcttccactg attcagtgag taactgttag gtggttccct
     2281 aagggattag gtatttcatc actgagctaa ccctggctat cattctgctt ttcttggctg
     2341 tctttcagat ttgactttat ttctaaaaat atttcaatgg gtcatatcac agattctttt
     2401 tttttaaatt aaagtaacat ttccaatcta ctaatgctaa tactgtttcg tatttatagc
     2461 tgatttgatg gagttggaca tggccatgga accagacaga aaagcggctg ttagtcactg
     2521 gcagcaacag tcttacctgg actctggaat ccattctggt gccactacca cagctccttc
     2581 tctgagtggt aaaggcaatc ctgaggaaga ggatgtggat acctcccaag tcctgtatga
     2641 gtgggaacag ggattttctc agtccttcac tcaagaacaa gtagctggta agagtattat
     2701 ttttcattgc cttactgaaa gtcagaatgc agttttgaga actaaaaagt tagtgtataa
     2761 tagtttaaat aaaatgttgt ggtgaagaaa agagagtaat agcaatgtca cttttaccat
     2821 ttaggatagc aaatacttag gtaaatgctg aactgtggat agtgagtgtt gaattaacct
     2881 tttccagata ttgatggaca gtatgcaatg actcgagctc agagggtacg agctgctatg
     2941 ttccctgaga cattagatga gggcatgcag atcccatcta cacagtttga tgctgctcat
     3001 cccacatccc aatggcgcgc cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3061 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3121 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3181 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3241 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3301 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3361 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3421 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3481 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3541 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3601 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3661 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3721 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3781 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3841 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3901 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3961 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4021 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4081 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4141 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4201 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4261 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4321 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4381 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4441 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4501 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4561 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4621 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4681 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4741 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4801 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4861 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4921 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4981 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5041 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5101 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5161 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5221 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5281 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5341 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5401 gcttaactat gcggcatcag agcagattgt actgagagtg cacca
//
