LOCUS       sequence-270852-        7917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Focal Adhesions, Excitation: 548, Emission: 561.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7917)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 7917)
  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: mKOet-Vinculin-23
FEATURES             Location/Qualifiers
     source          1..7917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     527..547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      615..624
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             648..1298
                     /codon_start=1
                     /product="monomeric Kusabira-Orange fluorescent protein 
                     (Karasawa et al., 2004)"
                     /label=mKO
                     /translation="MAVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     polyA_signal    4714..4835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4751..4770)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4805..4824
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4842..5297)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4979..5000)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5191..5210
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5324..5428
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5430..5787
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5430..5621
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5453..5473)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5638..5773
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5821..6615
                     /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             5821..6615
                     /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(5875..5894)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6485..6504
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6803..6822)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6847..6894
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7223..7811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7712..7731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      421 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      481 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      541 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      601 tagcgctacc ggtcgccacc atggtgagca agggcgagga gaataacatg gccgtgatta
      661 aaccagagat gaagatgagg tactacatgg acggctccgt caatgggcat gagttcacaa
      721 ttgaaggtga aggcacaggc agaccttacg agggacatca agagatgaca ctacgcgtca
      781 caatggccaa gggcgggcca atgcctttcg cgtttgactt agtgtcacac gtgttctgtt
      841 acggccacag accttttact aaatatccag aagagatacc agactatttc aaacaagcat
      901 ttcctgaagg cctgtcatgg gaaaggtcgt tggagttcga agatggtggg tccgcttcag
      961 tcagtgcgca tataagcctt agaggaaaca ccttctacca caaatccaaa tttactgggg
     1021 ttaactttcc tgccgatggt cctatcatgc aaaaccaaag tgttgattgg gagccatcaa
     1081 ccgagaaaat tactgccagc gacggagttc tgaagggtga tgttacgatg tacctaaaac
     1141 ttgaaggagg cggcaatcac aaatgccaat tcaagactac ttacaaggcg gcaaaaaaga
     1201 ttcttaaaat gccaggaagc cattacatca gccatcgcct cgtcaggaaa accgaaggca
     1261 acattactga gctggtagaa gatgcagtag ctcattccgg cggcatggac gagctgtaca
     1321 agtccggact cagatctcga gctcaagctt cgaattcggg aaactcgaga acaaagcttg
     1381 aaaacggccg catgccagtg tttcatacgc gcacgatcga gagcatcctg gagccggtgg
     1441 cacagcagat ctcccacctg gtgataatgc acgaggaggg cgaggtggac ggcaaagcca
     1501 ttcctgacct caccgcgccc gtggccgccg tgcaggcggc cgtcagcaac ctcgtccggg
     1561 ttggaaaaga gactgttcaa accactgagg atcagatttt gaagagagat atgccaccag
     1621 catttattaa ggttgagaat gcttgcacca agcttgtcca ggcagctcag atgcttcagt
     1681 cagaccctta ctcagtgcct gctcgagatt atctaattga tgggtcaagg ggcatcctct
     1741 ctggaacatc agacctgctc cttaccttcg atgaggctga ggtccgtaaa attattagag
     1801 tttgcaaagg aattttggaa tatcttacag tggcagaggt ggtggagact atggaagatt
     1861 tggtcactta cacaaagaat cttgggccag gaatgactaa gatggccaag atgattgacg
     1921 agagacagca ggagctcact caccaggagc accgagtgat gttggtgaac tcgatgaaca
     1981 ccgtgaaaga gttgctgcca gttctcattt cagctatgaa gatttttgta acaactaaaa
     2041 actcaaaaaa ccaaggcata gaggaagctt taaaaaatcg caattttact ctagaaaaaa
     2101 tgagtgctga aattaatgag ataattcgtg tgttacaact cacctcttgg gatgaagatg
     2161 cctgggccag caaggacact gaagccatga agagagcatt ggcctccata gactccaaac
     2221 tgaaccaggc caaaggttgg ctccgtgacc ctagtgcctc cccaggggat gctggtgagc
     2281 aggccatcag acagatctta gatgaagctg gaaaagttgg tgaactctgt gcaggcaaag
     2341 aacgcaggga gattctggga acttgcaaaa tgctagggca gatgactgat caagtggctg
     2401 acctccgtgc cagaggacaa ggatcctcac cggtggccat gcagaaagct cagcaggtat
     2461 ctcagggtct ggatgtgctc acagcaaaag tggaaaatgc agctcgcaag ctggaagcca
     2521 tgaccaactc aaagcagagc attgcaaaga agatcgatgc tgctcagaac tggcttgcag
     2581 atccaaatgg tggaccggaa ggagaagagc agattcgagg tgctttggct gaagctcgga
     2641 aaatagcaga attatgtgat gatcctaaag aaagagatga cattctacgt tcccttgggg
     2701 aaatatctgc tctgacttct aaattagcag atctacgaag acaggggaaa ggagattctc
     2761 cagaggctcg agccttggcc aaacaggtgg ccacggccct gcagaacctg cagaccaaaa
     2821 ccaaccgggc tgtggccaac agcagaccgg ccaaagcagc tgtacacctt gagggcaaga
     2881 ttgagcaagc acagcggtgg attgataatc ccacagtgga tgaccgtgga gtcggtcagg
     2941 ctgccatccg ggggcttgtg gccgaagggc atcgtctggc taatgttatg atggggcctt
     3001 atcggcaaga tcttctcgcc aagtgtgacc gagtggacca gctgacagcc cagctggctg
     3061 acctggctgc cagaggggaa ggggagagtc ctcaggcacg agcacttgca tctcagctcc
     3121 aagactcctt aaaggatcta aaagctcgga tgcaggaggc catgactcag gaagtgtcag
     3181 atgttttcag cgataccaca actcccatca agctgttggc agtggcagcc acggcgcctc
     3241 ctgatgcgcc taacagggaa gaggtatttg atgagagggc agctaacttt gaaaaccatt
     3301 caggaaagct tggtgctacg gccgagaagg cggctgcggt tggtactgct aataaatcaa
     3361 cagtggaagg cattcaggcc tcagtgaaga cggcccgaga actcacaccc caggtggtct
     3421 cggctgctcg tatcttactt aggaaccctg gaaatcaagc tgcttatgaa cattttgaga
     3481 ccatgaagaa ccagtggatc gataatgttg aaaaaatgac agggctggtg gacgaagcca
     3541 ttgataccaa atctctgttg gatgcttcag aagaagcaat taaaaaagac ctggacaagt
     3601 gcaaggtagc tatggccaac attcagcctc agatgctggt tgctggggca accagtattg
     3661 ctcgtcgggc caaccggatc ctgctggtgg ctaagaggga ggtggaaaat tccgaggatc
     3721 ccaagttccg tgaggctgtg aaagctgcct ctgatgaatt gagcaaaacc atctccccga
     3781 tggtgatgga tgcaaaagct gtggctggaa acatttccga ccctggactg caaaagagct
     3841 tcctggactc aggatatcgg atcctgggag ctgtggccaa ggtcagagaa gccttccaac
     3901 ctcaggagcc tgacttcccg ccgcctccac cagaccttga acaactccga ctaacagatg
     3961 agcttgctcc tcccaaacca cctctgcctg aaggtgaggt ccctccacct aggcctccac
     4021 caccagagga aaaggatgaa gagttccctg agcagaaggc cggggaggtg attaaccagc
     4081 caatgatgat ggctgccaga cagctccatg atgaagctcg caaatggtcc agcaagggca
     4141 atgacatcat tgcagcagcc aagcgcatgg ctctgctgat ggctgagatg tctcggctgg
     4201 taagaggggg cagtggtacc aagcgggcac tcattcagtg tgccaaggac atcgccaagg
     4261 cctcagatga ggtgactcgg ttggccaagg aggttgccaa gcagtgcaca gataaacgga
     4321 ttagaaccaa cctcttacag gtatgtgagc gaatcccaac cataagcacc cagctcaaaa
     4381 tcctgtccac agtgaaggcc accatgctgg gccggaccaa catcagtgat gaggagtctg
     4441 agcaggccac agagatgctg gttcacaatg cccagaacct catgcagtct gtgaaggaga
     4501 ctgtgcggga agctgaagct gcttcaatca aaattcgaac agatgctgga tttacactgc
     4561 gctgggttag aaagactccc tggtaccagt aggcggccgc gactctagat cataatcagc
     4621 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     4681 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     4741 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4801 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     4861 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4921 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4981 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     5041 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     5101 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     5161 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     5221 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     5281 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5341 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5401 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     5461 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5521 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5581 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5641 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5701 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5761 ttttttggag gcctaggctt ttgcaaagat catcaagaga caggatgagg atcgtttcgc
     5821 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5881 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5941 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6001 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6061 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6121 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6181 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6241 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6301 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     6361 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6421 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6481 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6541 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6601 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6661 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6721 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6781 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     6841 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     6901 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     6961 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     7021 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     7081 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7141 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7201 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7261 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7321 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7381 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7441 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7501 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7561 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7621 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7681 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7741 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7801 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7861 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccg
//
