LOCUS       sequence-337282-        6433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Endoplasmic Reticulum, Excitation: 551, Emission: 565.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6433)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 6433)
  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-Calnexin-N-14
FEATURES             Location/Qualifiers
     source          1..6433
                     /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      2377..2386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2383..3039
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     polyA_signal    3161..3282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3198..3217)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3252..3271
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3289..3744)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3426..3447)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3638..3657
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3771..3875
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3877..4234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3877..4068
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3900..3920)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4085..4220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4269..5063
                     /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             4269..5063
                     /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(4323..4342)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4933..4952
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5251..5270)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5295..5342
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5671..6259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6160..6179
                     /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 ccggactcag atctcgagac cgccatggaa gggaagtggt tgctgtgtat gttactggtg
      601 cttggaactg ctattgttga ggctcatgat ggacatgatg atgatgtgat tgatattgag
      661 gatgaccttg acgatgtcat tgaagaggta gaagactcaa aaccagatac cactgctcct
      721 ccttcatctc ccaaggttac ttacaaagct ccagttccaa caggggaagt atattttgct
      781 gattcttttg acagaggaac tctgtcaggg tggattttat ccaaagccaa gaaagacgat
      841 accgatgatg aaattgccaa atatgatgga aagtgggagg tagaggaaat gaaggagtca
      901 aagcttccag gtgataaagg acttgtgttg atgtctcggg ccaagcatca tgccatctct
      961 gctaaactga acaagccctt cctgtttgac accaagcctc tcattgttca gtatgaggtt
     1021 aatttccaaa atggaataga atgtggtggt gcctatgtga aactgctttc taaaacacca
     1081 gaactcaacc tggatcagtt ccatgacaag accccttata cgattatgtt tggtccagat
     1141 aaatgtggag aggactataa actgcacttc atcttccgac acaaaaaccc caaaacgggt
     1201 atctatgaag aaaaacatgc taagaggcca gatgcagatc tgaagaccta ttttactgat
     1261 aagaaaacac atctttacac actaatcttg aatccagata atagttttga aatactggtt
     1321 gaccaatctg tggtgaatag tggaaatctg ctcaatgaca tgactcctcc tgtaaatcct
     1381 tcacgtgaaa ttgaggaccc agaagaccgg aagcccgagg attgggatga aagaccaaaa
     1441 atcccagatc cagaagctgt caagccagat gactgggatg aagatgcccc tgctaagatt
     1501 ccagatgaag aggccacaaa acccgaaggc tggttagatg atgagcctga gtacgtacct
     1561 gatccagacg cagagaaacc tgaggattgg gatgaagaca tggatggaga atgggaggct
     1621 cctcagattg ccaaccctag atgtgagtca gctcctggat gtggtgtctg gcagcgacct
     1681 gtgattgaca accccaatta taaaggcaaa tggaagcctc ctatgattga caatcccagt
     1741 taccagggaa tctggaaacc caggaaaata ccaaatccag atttctttga agatctggaa
     1801 cctttcagaa tgactccttt tagtgctatt ggtttggagc tgtggtccat gacctctgac
     1861 attttttttg acaactttat catttgtgct gatcgaagaa tagttgatga ttgggccaat
     1921 gatggatggg gcctgaagaa agctgctgat ggggctgctg agccaggcgt tgtggggcag
     1981 atgaacgagg cagctgaaga gcgcccgtgg ctgtgggtag tctatattct aactgtagcc
     2041 cttcctgtgt tcctggttat cctcttctgc tgttctggaa agaaacagac cagtggtatg
     2101 gagtataaga aaactgatgc acctcaaccg gatgtgaagg aagaggaaga agagaaggaa
     2161 gaggaaaagg acaagggaga tgaggaggag gaaggagaag agaaacttga agagaaacag
     2221 aaaagtgatg ctgaagaaga tggtggcact gtcagtcaag aggaggaaga cagaaaacct
     2281 aaagcagagg aggatgaaat tttgaacaga tcaccaagaa acagaaagcc acgaagagag
     2341 gggggtgcgt caggcggcag cggatctgga ccggtcgcca ccatggtgag tgtgattaaa
     2401 ccagagatga agatgaggta ctacatggac ggctccgtca atgggcatga gttcacaatt
     2461 gaaggtgaag gcacaggcag accttacgag ggacatcaag agatgacact acgcgtcaca
     2521 atggccgagg gcgggccaat gcctttcgcg tttgacttag tgtcacacgt gttctgttac
     2581 ggccacagag tatttactaa atatccagaa gagataccag actatttcaa acaagcattt
     2641 cctgaaggcc tgtcatggga aaggtcgttg gagttcgaag atggtgggtc cgcttcagtc
     2701 agtgcgcata taagccttag aggaaacacc ttctaccaca aatccaaatt tactggggtt
     2761 aactttcctg ccgatggtcc tatcatgcaa aaccaaagtg ttgattggga gccatcaacc
     2821 gagaaaatta ctgccagcga cggagttctg aagggtgatg ttacgatgta cctaaaactt
     2881 gaaggaggcg gcaatcacaa atgccaaatg aagactactt acaaggcggc aaaagagatt
     2941 cttgaaatgc caggagacca ttacatcggc catcgcctcg tcaggaaaac cgaaggcaac
     3001 attactgagc aggtagaaga tgcagtagct catagctaag cggccgcgac tctagatcat
     3061 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3121 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3181 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3241 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3301 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3361 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3421 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3481 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3541 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3601 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3661 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3721 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3781 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3841 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3901 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4021 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4081 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4141 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4201 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4261 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4321 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4381 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4441 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4501 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4561 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4621 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4681 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4741 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4801 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4861 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4921 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4981 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5041 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5101 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5161 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5221 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5281 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5341 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5401 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5461 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5521 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5581 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5641 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5701 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5761 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5821 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5881 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5941 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6001 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6061 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6121 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6181 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6241 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6301 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6361 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     6421 atatggagtt ccg
//
