LOCUS       sequence-485363-        5650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and LE-mEGFP sequence for C-terminus tagging of human 
            cadherin 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5650)
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5650)
  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            "Unpublished"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-166:CDH1-mEGFP
FEATURES             Location/Qualifiers
     source          1..5650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     protein_bind    148..168
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             join(1180..1182,1183..1896)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1222..1243)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1483..1502)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1830..1851
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     repeat_region   2913..3053
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2913..3042
                     /label=AAV2 ITR
     rep_origin      3128..3583
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3215..3234)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3425..3446
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3600..3619)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3719..3741
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3779..3797)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3865..3969
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3970..4038,4039..4830)
                     /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(4188..4207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5001..5589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5490..5509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttctagacaa ctttgtatag aaaagttggg tacctacttt
      181 atttatttca taaaatgaaa atatataaag aaaaggccgg gtgcagtggc tcacgcctgt
      241 aatcccagca ctttgggagg ccgaggcagg tggatcacct gagatcagga gttagagacc
      301 agcctggcca acatggtgaa accccgtttt tactaaaaat acaaaaatta gccgggcatg
      361 gtggcgcatg cctgtagacc cagctacttg ggaggctgag gcaggagaat cgcttgaacc
      421 tgggaggcag atgttgcagt gagccaaaat cacaccattg cactccagcc tgtgtgacaa
      481 gagtgaaacc ctgtctctaa aaaaaaagaa aaagaaaaaa aatacaagtc cttccttagt
      541 tcccacaccc ccactcccca gatttaactg ttaagcattt ggcatttatc cttccaggcc
      601 tttctgggtg gaaatgcagc ctgcatgcac cagtatgcaa agcttccagc tctgcccatc
      661 ccagctggtg gtgtggcagc cccactctga tctatgggga ctccatcctg actggttggt
      721 gcttgcatct ctcctgccta catataactc accatttcac tttctctttc catcaacaac
      781 atatcctgtg cttctgcatc cttccatgac agtgtgtata aatgtacgtt gttggtgttc
      841 actgctccgt ggtgtgccac aagtctgggt gcattgtcgt accttacata ttgctagact
      901 tcttgcccca gatgacaggt gtgcccttcc tttcactaaa agatgctttt gtcccttctt
      961 ctttagaatc tgaaagcggc tgatactgac cccacagccc cgccttatga ttctctgctc
     1021 gtgtttgact atgaaggaag cggttccgaa gctgctagtc tgagctccct gaactcctca
     1081 gagtcagaca aagaccagga ctatgactac ttgaacgaat ggggcaatcg cttcaagaag
     1141 ctggctgaca tgtacggagg cggcgaggac gaccttgaag tgagcaaggg cgaggagctg
     1201 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1261 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1321 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1381 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1441 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1501 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1561 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1621 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1681 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1741 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
     1801 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1861 gggatcactc tcggcatgga cgagctgtac aagtagggga ctcgagagac gcgggcgcca
     1921 gacccatgtg cttggaaatg cagaaatcac gttgctggtg gtttttcagc tcccttccct
     1981 tgagatgagt ttctggggaa aaaaaagaga ctggttagtg atgcagttag tatagcttta
     2041 tactctctcc actttatagc tctaataagt ttgtgttaga aaagtttcga cttatttctt
     2101 aaagcttttt tttttttccc atcactcttt acatggtggt gatgtccaaa agatacccaa
     2161 attttaatat tccagaagaa caactttagc atcagaaggt tcacccagca ccttgcagat
     2221 tttcttaagg aattttgtct cacttttaaa aagaagggga gaagtcagct actctagttc
     2281 tgttgttttg tgtatataat tttttaaaaa aaatttgtgt gcttctgctc attactacac
     2341 tggtgtgtcc ctctgccttt tttttttttt taagacaggg tctcattcta tcggccaggc
     2401 tggagtgcag tggtgcaatc acagctcact gcagccttgt cctcccaggc tcaagctatc
     2461 cttgcacctc agcctcccaa gtagctggga ccacaggcat gcaccactac gcatgactaa
     2521 ttttttaaat atttgagacg gggtctccct gtgttaccca ggctggtctc aaactcctgg
     2581 gctcaagtga tcctcccatc ttggcctccc agagtattgg gattacagac atgagccact
     2641 gcacctgccc agctccccaa ctccctgcca ttttttaaga gacagtttcg ctccatcgcc
     2701 caggcctggg atgcagtgat gtgatcatag ctcactgtaa cctcaaactc tggggctcaa
     2761 gcagttctcc caccagcctc ctttttattt ttttgtacag atggggtctt gctatgttgc
     2821 ccaagctggt cttaaactcc tggcctcaag caatccttct gccttggccc cccaaagtgc
     2881 tgggattgtg ggcgcatgct ggggagggcc gcaggaaccc ctagtgatgg agttggccac
     2941 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3001 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     3061 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3121 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3181 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     3241 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3301 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3361 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3421 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     3481 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     3541 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3601 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3661 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3721 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3781 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3841 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3901 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3961 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4021 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4081 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4141 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4201 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4261 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4321 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4381 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4441 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4501 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4561 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4621 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4681 cgtggaagcc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4741 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4801 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4861 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4921 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4981 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5041 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5101 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5161 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5221 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5281 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5341 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5401 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5461 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5521 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5581 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5641 gctcacatgt
//
