LOCUS       sequence-177099-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Tight Junctions, Excitation: 433, Emission: 475.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  TITLE     Davidson Cyan Fluorescent Proteins
REFERENCE   2  (bases 1 to 7083)
  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"
COMMENT     Sequence Label: mCerulean-Beta-Catenin-20
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1324..1333
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1375..1410
                     /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"
     polyA_signal    3811..3932
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3848..3867)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3902..3921
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3939..4394)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4076..4097)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4288..4307
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4421..4525
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4527..4884
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4527..4718
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4550..4570)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4735..4870
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4919..5713
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4919..5713
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4973..4992)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5583..5602
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5901..5920)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5945..5992
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6321..6909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6810..6829
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcgc ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaacgcc atcagcgaca acgtctatat caccgccgac
     1021 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1321 gcggccacca tggctactca agctgacctg atggagttgg acatggccat ggagccggac
     1381 agaaaagctg ctgtcagcca ctggcagcag cagtcttact tggattctgg aatccattct
     1441 ggtgccacca ccacagctcc ttccctgagt ggcaagggca accctgagga agaagatgtt
     1501 gacacctccc aagtccttta tgaatgggag caaggctttt cccagtcctt cacgcaagag
     1561 caagtagctg atattgacgg gcagtatgca atgactaggg ctcagagggt ccgagctgcc
     1621 atgttccctg agacgctaga tgagggcatg cagatcccat ccacgcagtt tgacgctgct
     1681 catcccacta atgtccagcg cttggctgaa ccatcacaga tgttgaaaca tgcagttgtc
     1741 aatttgatta actatcagga tgacgcggaa cttgccacac gtgcaattcc tgagctgaca
     1801 aaactgctaa acgatgagga ccaggtggta gttaataaag ctgctgttat ggtccatcag
     1861 ctttccaaaa aggaagcttc cagacatgcc atcatgcgct cccctcagat ggtgtctgcc
     1921 attgtacgca ccatgcagaa tacaaatgat gtagagacag ctcgttgtac tgctgggact
     1981 ctgcacaacc tttctcacca ccgcgagggc ttgctggcca tctttaagtc tggtggcatc
     2041 ccagcgctgg tgaaaatgct tgggtcacca gtggattctg tactgttcta cgccatcacg
     2101 acactgcata atctcctgct ccatcaggaa ggagctaaaa tggcagtgcg cctagctggt
     2161 ggactgcaga aaatggttgc tttgctcaac aaaacaaacg tgaaattctt ggctattaca
     2221 acagactgcc ttcagatctt agcttatggc aatcaagaga gcaagctcat cattctggcc
     2281 agtggtggac cccaagcctt agtaaacata atgaggacct acacttatga gaagcttctg
     2341 tggaccacaa gcagagtgct gaaagtgctg tctgtctgct ctagcaacaa gccggccatt
     2401 gtagaagctg gtgggatgca ggcactgggg cttcatctga cagacccaag tcagcgactt
     2461 gttcaaaact gtctttggac tctcagaaac ctttcagatg cagcgactaa gcaggaaggg
     2521 atggaaggcc tccttgggac tctagtgcag cttctgggtt ccgatgatat aaatgtggtc
     2581 acctgtgcag ctggaattct ctctaacctc acttgcaata attacaaaaa caagatgatg
     2641 gtgtgccaag tgggtggcat agaggctctt gtacgcaccg tccttcgtgc tggtgacagg
     2701 gaagacatca ctgagcctgc catctgtgct cttcgtcatc tgaccagccg gcatcaggaa
     2761 gccgagatgg cccagaatgc cgttcgcctt cattatggac tgcctgttgt ggttaaactc
     2821 ctgcacccac catcccactg gcctctgata aaggcaactg ttggattgat tcgaaacctt
     2881 gccctttgcc cagcaaatca tgcgcctttg cgggaacagg gtgctattcc acgactagtt
     2941 cagctgcttg tacgagcaca tcaggacacc caacggcgca cctccatggg tggaacgcag
     3001 cagcagtttg tggagggcgt gcgcatggag gagatagtag aagggtgtac tggagctctc
     3061 cacatccttg ctcgggacgt tcacaaccgg attgtaatcc gaggactcaa taccattcca
     3121 ttgtttgtgc agttgcttta ttctcccatt gaaaatatcc aaagagtagc tgcaggggtc
     3181 ctctgtgaac ttgctcagga caaggaggct gcagaggcca ttgaagctga gggagccaca
     3241 gctcccctga cagagttact ccactccagg aatgaaggcg tggcaacata cgcagctgct
     3301 gtcctattcc gaatgtctga ggacaagcca caggattaca agaagcggct ttcagtcgag
     3361 ctgaccagtt ccctcttcag gacagagcca atggcttgga atgagactgc agatcttgga
     3421 ctggacattg gtgcccaggg agaagccctt ggatatcgcc aggatgatcc cagctaccgt
     3481 tcttttcact ctggtggata cggccaggat gccttgggga tggaccctat gatggagcat
     3541 gagatgggtg gccaccaccc tggtgctgac tatccagttg atgggctgcc tgatctggga
     3601 cacgcccagg acctcatgga tgggctgccc ccaggtgata gcaatcagct ggcctggttt
     3661 gatactgacc tgtaaaagct tggatccacc ggatctagat aactgatcat aatcagccat
     3721 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3781 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3841 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3901 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     3961 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4021 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4081 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4141 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4201 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4261 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4321 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4381 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4441 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4501 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4561 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4621 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4681 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4741 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4801 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4861 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4921 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4981 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5041 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5101 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5161 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5221 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5281 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5341 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5401 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5461 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5521 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5581 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5641 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5701 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5761 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5821 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5881 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5941 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     6001 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6061 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6121 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     6181 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6241 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6301 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6361 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6421 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6481 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6541 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6601 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6661 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6721 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6781 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6841 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6901 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6961 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     7021 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     7081 ccg
//
