LOCUS       sequence-210571-        9858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Tight Junctions, Excitation: 551, Emission: 565.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9858)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 9858)
  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: mKO2-ZO1-C-14
FEATURES             Location/Qualifiers
     source          1..9858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             671..1324
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     RBS             3025..3033
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    6704..6825
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6741..6760)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6795..6814
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6832..7287)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6969..6990)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7181..7200
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7314..7418
                     /gene="bla"
                     /label=AmpR promoter
     promoter        7420..7777
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7420..7611
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7443..7463)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7628..7763
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7812..8606
                     /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             7812..8606
                     /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(7866..7885)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8476..8495
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8794..8813)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8838..8885
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      9214..9802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9703..9722
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggtcgccacc atggtgagtg tgattaaacc agagatgaag atgaggtact acatggacgg
      721 ctccgtcaat gggcatgagt tcacaattga aggtgaaggc acaggcagac cttacgaggg
      781 acatcaagag atgacactac gcgtcacaat ggccgagggc gggccaatgc ctttcgcgtt
      841 tgacttagtg tcacacgtgt tctgttacgg ccacagagta tttactaaat atccagaaga
      901 gataccagac tatttcaaac aagcatttcc tgaaggcctg tcatgggaaa ggtcgttgga
      961 gttcgaagat ggtgggtccg cttcagtcag tgcgcatata agccttagag gaaacacctt
     1021 ctaccacaaa tccaaattta ctggggttaa ctttcctgcc gatggtccta tcatgcaaaa
     1081 ccaaagtgtt gattgggagc catcaaccga gaaaattact gccagcgacg gagttctgaa
     1141 gggtgatgtt acgatgtacc taaaacttga aggaggcggc aatcacaaat gccaaatgaa
     1201 gactacttac aaggcggcaa aagagattct tgaaatgcca ggagaccatt acatcggcca
     1261 tcgcctcgtc aggaaaaccg aaggcaacat tactgagcag gtagaagatg cagtagctca
     1321 tagctccgga agtgctggga gcgcaagcgg aagcggaggc tccggtgagg aaacagctat
     1381 atgggaacaa catacagtga cgcttcacag ggctcctgga tttggatttg gaattgcaat
     1441 atctggtgga cgagataatc ctcattttca gagtggggaa acgtcaatag tgatttcaga
     1501 tgtgctgaaa ggaggaccag ctgaaggaca gctacaggaa aatgaccgag ttgcaatggt
     1561 taacggagtt tcaatggata atgttgaaca tgcttttgct gttcagcaac taaggaaaag
     1621 tgggaaaaat gcaaaaatta caattagaag gaagaagaaa gttcaaatac cagtaagtcg
     1681 tcctgatcct gaaccagtat ctgataatga agaagatagt tatgatgagg aaatacatga
     1741 tccaagaagt ggccggagtg gtgtggttaa cagaaggagt gagaagattt ggccgaggga
     1801 tagaagtgca agtagagaga ggagcttgtc cccgcggtca gacaggcggt cagtggcttc
     1861 cagccagcct gctaaaccta ctaaagtcac actggtgaaa tcccggaaaa atgaagaata
     1921 tggtcttcga ttggcaagcc atatatttgt taaggaaatt tcacaagata gtttggcagc
     1981 aagagatggc aatattcaag aaggtgatgt tgtattgaag ataaatggta ctgtgacaga
     2041 aaatatgtca ttgacagatg caaagacatt gatagaaagg tctaaaggca aattaaaaat
     2101 ggtagttcaa agagatgaac gggctacgct attgaatgtc cctgatcttt ctgacagcat
     2161 ccactctgct aatgcctctg agagagacga catttcagaa attcagtcac tggcatcaga
     2221 tcattctggt cgatcacacg ataggcctcc ccgccgcagc cggtcacgat ctcctgacca
     2281 gcggtcagag ccttctgatc attccaggca ctcgccgcag cagccaagca atggcagtct
     2341 gcggagtaga gatgaagaga gaatttctaa acctggggct gtctcaactc ctgtaaagca
     2401 tgctgatgat cacacaccta aaacagtgga agaagttaca gttgaaagaa atgagaaaca
     2461 aacaccttct cttccagaac caaagcctgt gtatgcccaa gttggcaacc agatgtggat
     2521 ttacctgtca gtccatctga tggtgtccta cctaattcaa ctcatgaaga tgggatttct
     2581 tcggcccagc atgaaattgg taaaattcag aaaaggagat agtgtgggtt tgcggctggc
     2641 tggtggaaat gatgttggaa tatttgtagc tggcgttcta gaagatagcc ctgcagccaa
     2701 ggaaggctta gaggaaggtg atcaaattct cagggtaaac aacgtagatt ttacaaatat
     2761 cataagagaa gaagccgtcc ttttcctgct tgacctccct aaaggagaag aagtgaccat
     2821 attggctcag aagaagaagg atgtttatcg tcgcattgta gaatcagatg taggagattc
     2881 tttctatatt agaacccatt ttgaatatga aaaggaatct ccctatggac ttagttttaa
     2941 caaaggagag gtgttccgtg ctgtggatac cttgtacaat ggaaaactgg gctcttggct
     3001 tgctattcga attggtaaaa atcataagga ggtagaacga ggcatcatcc ctaataagaa
     3061 cagagctgag cagctcgcca gtgtacagta tacacttcca aaaacagcag gcggagaccg
     3121 tgctgacttc tggagattca gaggtcttcg cagctccaag agaaatcttc gaaaaagcag
     3181 agaggatttg tccgctcagc ctgttcaaac aaagtttcca gcttatgaaa gagtggttct
     3241 tcgagaagct ggatttctga ggcctgtaac catttttgga ccaatagctg atgttgccag
     3301 agaaaagctg gcaagagaag aaccagatat ttatcaaatt gcaaagagtg aaccacgaga
     3361 cgctggaact gaccaacgta gctctggcta tattcgcctg catacaataa agcaaatcat
     3421 agatcaagac aaacatgctt tattagatgt aacaccaaat gcagttgatc gtcttaacta
     3481 tgcccagtgg tatccaattg ttgtatttct taaccctgat tctaagcaag gagtaaaaac
     3541 aatgagaatg aggttatgtc cagaatctcg gaaaagtgcc aggaagttat acgagcgatc
     3601 tcataaactt gctaaaaata atcaccatct ttttacaact acaattaact taaattcaat
     3661 gaatgatggt tggtatggtg cgctgaaaga agcagttcaa caacagcaaa accagctggt
     3721 atgggtttcc gagggaaagg cggatggtgc tacaagtgat gaccttgatt tgcatgatga
     3781 tcgtctgtcc tacctgtcag ctccaggtag tgaatactca atgtatagca cggacagtag
     3841 acacacttct gactatgaag acacagacac agaaggcggg gcctacactg atcaagaact
     3901 agatgaaact cttaatgatg aggttgggac tccaccggag tctgccatta cacggtcctc
     3961 tgagcctgta agagaggact cctctggaat gcatcatgaa aaccaaacat atcctcctta
     4021 ctcaccacaa gcgcagccac aaccaattca tagaatagac tcccctggat ttaagccagc
     4081 ctctcaacag aaagcagaag cttcatctcc agtcccttac ctttcgcctg aaacaaaccc
     4141 agcatcatca acctctgctg ttaatcataa tgtaaattta actaatgtca gactggagga
     4201 gcccacccca gctccttcca cctcttactc accacaagct gattctttaa gaacaccaag
     4261 tactgaggca gctcacataa tgctaagaga tcaagaacca tcattgtcgt cgcatgtaga
     4321 tccaacaaag gtgtatagaa aggacccata tcccgaggaa atgatgaggc agaaccatgt
     4381 tttgaaacag ccagccgtta gtcacccagg gcacaggcca gacaaagagc ctaatctgac
     4441 ctatgaaccc caactcccat acgtagagaa acaagccagc agagacctgg agcagcccac
     4501 atacagatac gagtcctcaa gctatacgga ccagttttct cgaaactatg aacatcgtct
     4561 gcgatacgaa gatcgcgtcc ccatgtatga agaacagtgg tcatattatg atgacaaaca
     4621 gccctaccca tctcggccac cttttgataa tcagcactct caagaccttg actccagaca
     4681 gcatcccgaa gagtcctcag aacgagggta ctttccacgt tttgaagagc cagcccctct
     4741 gtcttacgac agcagaccac gttacgaaca ggcacctaga gcatccgccc tgcggcacga
     4801 agagcagcca gctcctgggt atgacacaca tggtagactc agaccggaag cccagcccca
     4861 cccttcagca gggcccaagc ctgcagagtc caagcagtat tttgagcaat attcacgcag
     4921 ttacgagcaa gtaccacccc aaggatttac ctctagagca ggtcattttg agcctctcca
     4981 tggtgctgca gctgtccctc cgctgatacc ttcatctcag cataagccag aagctctgcc
     5041 ttcaaacacc aaaccactgc ctccaccccc aactcaaacc gaagaagagg aagatccagc
     5101 aatgaagcca cagtctgtac tcaccagagt taagatgttt gaaaacaaaa ggtctgcatc
     5161 cttagagacc aagaaggatg taaatgacac tggcagtttt aagcctccag aagtagcatc
     5221 taaaccttca ggtgctccca tcattggtcc caaacccact tctcagaatc aattcagtga
     5281 acatgacaaa actctgtaca ggattccaga acctcaaaaa cctcaactga agccacctga
     5341 agatattgtt cggtccaatc attatgaccc tgaagaagat gaagaatatt atcgaaaaca
     5401 gctgtcatac tttgaccgaa gaagttttga gaataagcct cctgcacaca ttgctgccag
     5461 ccatctctcc gagcctgcaa agccagctca ttctcagaat caatcaaatt tttctagtta
     5521 ttcttcaaag ggaaagcctc ctgaagctga tggtgtggat agatcatttg gcgagaaacg
     5581 ctatgaaccc atccaggcca ctccccctcc tcctccattg ccctcgcagt atgcccagcc
     5641 atctcagcct gtcaccagcg cgtctctcca catacattct aagggagcac atggtgaagg
     5701 taattcagtg tcattggatt ttcagaattc cttagtgtcc aaaccagacc cacctccatc
     5761 tcagaataag ccagcaactt tcagaccacc aaaccgagaa gatactgctc aggcagcttt
     5821 ctatccccag aaaagttttc cagataaagc cccagttaat ggaactgaac agactcagaa
     5881 aacagtcact ccagcataca atcgattcac accaaaacca tatacaagtt ctgcccgacc
     5941 atttgaacgc aagtttgaaa gtcctaaatt caatcacaat cttctgccaa gtgaaactgc
     6001 acataaacct gacttgtctt caaaaactcc cacttctcca aaaactcttg tgaaatcgca
     6061 cagtttggca cagcctcctg agtttgacag tggagttgaa actttctcta tccatgcaga
     6121 gaagcctaaa tatcaaataa ataatatcag cacagtgcct aaagctattc ctgtgagtcc
     6181 ttcagctgtg gaagaggatg aagatgaaga tggtcatact gtggtggcca cagcccgagg
     6241 catatttaac agcaatgggg gcgtgctgag ttccatagaa actggtgtta gtataattat
     6301 ccctcaagga gccattcccg aaggagttga gcaggaaatc tatttcaagg tctgccggga
     6361 caacagcatc cttccacctt tagataaaga gaaaggtgaa acactgctga gtcctttggt
     6421 gatgtgtggt ccccatggcc tcaagttcct gaagcctgtg gagctgcgct taccacactg
     6481 tgatcctaaa acctggcaaa acaagtgtct tcccggagat ccaaattatc tcgttggagc
     6541 aaactgtgtt tctgtcctta ttgaccactt ttaaggatcc accggatcta gataactgat
     6601 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     6661 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     6721 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6781 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     6841 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     6901 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     6961 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     7021 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     7081 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     7141 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     7201 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     7261 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     7321 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     7381 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     7441 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     7501 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     7561 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     7621 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     7681 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     7741 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     7801 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     7861 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     7921 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     7981 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     8041 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     8101 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     8161 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     8221 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     8281 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     8341 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     8401 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     8461 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     8521 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     8581 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     8641 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     8701 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     8761 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     8821 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     8881 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     8941 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     9001 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     9061 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     9121 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     9181 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     9241 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     9301 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     9361 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     9421 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     9481 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     9541 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     9601 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     9661 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     9721 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     9781 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     9841 ttgctggcct tttgctca
//
