LOCUS       sequence-218287-        6450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6450)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 6450)
  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-PLK1-N-16
FEATURES             Location/Qualifiers
     source          1..6450
                     /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      2394..2403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2400..3056
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     polyA_signal    3178..3299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3215..3234)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3269..3288
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3306..3761)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3443..3464)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3655..3674
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3788..3892
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3894..4251
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3894..4085
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3917..3937)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4102..4237
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4286..5080
                     /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             4286..5080
                     /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(4340..4359)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4950..4969
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5268..5287)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5312..5359
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5688..6276
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6177..6196
                     /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 gctagcgcca
      541 ccatgagtgc tgcagtgact gcagggaagc tggcacgggc accggccgac cctgggaaag
      601 ccggggtccc cggagttgca gctcccggag ctccggcggc ggctccaccg gcgaaagaga
      661 tcccggaggt cctagtggac ccacgcagcc ggcggcgcta tgtgcggggc cgctttttgg
      721 gcaagggcgg ctttgccaag tgcttcgaga tctcggacgc ggacaccaag gaggtgttcg
      781 cgggcaagat tgtgcctaag tctctgctgc tcaagccgca ccagagggag aagatgtcca
      841 tggaaatatc cattcaccgc agcctcgccc accagcacgt cgtaggattc cacggctttt
      901 tcgaggacaa cgacttcgtg ttcgtggtgt tggagctctg ccgccggagg tctctcctgg
      961 agctgcacaa gaggaggaaa gccctgactg agcctgaggc ccgatactac ctacggcaaa
     1021 ttgtgcttgg ctgccagtac ctgcaccgaa accgagttat tcatcgagac ctcaagctgg
     1081 gcaacctttt cctgaatgaa gatctggagg tgaaaatagg ggattttgga ctggcaacca
     1141 aagtcgaata tgacggggag aggaagaaga ccctgtgtgg gactcctaat tacatagctc
     1201 ccgaggtgct gagcaagaaa gggcacagtt tcgaggtgga tgtgtggtcc attgggtgta
     1261 tcatgtatac cttgttagtg ggcaaaccac cttttgagac ttcttgccta aaagagacct
     1321 acctccggat caagaagaat gaatacagta ttcccaagca catcaacccc gtggccgcct
     1381 ccctcatcca gaagatgctt cagacagatc ccactgcccg cccaaccatt aacgagctgc
     1441 ttaatgacga gttctttact tctggctata tccctgcccg tctccccatc acctgcctga
     1501 ccattccacc aaggttttcg attgctccca gcagcctgga ccccagcaac cggaagcccc
     1561 tcacagtcct caataaaggc ttggagaacc ccctgcctga gcgtccccgg gaaaaagaag
     1621 aaccagtggt tcgagagaca ggtgaggtgg tcgactgcca cctcagtgac atgctgcagc
     1681 agctgcacag tgtcaatgcc tccaagccct cggagcgtgg gctggtcagg caagaggagg
     1741 ctgaggatcc tgcctgcatc cccatcttct gggtcagcaa gtgggtggac tattcggaca
     1801 agtacggcct tgggtatcag ctctgtgata acagcgtggg ggtgctcttc aatgactcaa
     1861 cacgcctcat cctctacaat gatggtgaca gcctgcagta catagagcgt gacggcactg
     1921 agtcctacct caccgtgagt tcccatccca actccttgat gaagaagatc accctcctta
     1981 aatatttccg caattacatg agcgagcact tgctgaaggc aggtgccaac atcacgccgc
     2041 gcgaaggtga tgagctcgcc cggctgccct acctacggac ctggttccgc acccgcagcg
     2101 ccatcatcct gcacctcagc aacggcagcg tgcagatcaa cttcttccag gatcacacca
     2161 agctcatctt gtgcccactg atggcagccg tgacctacat cgacgagaag cgggacttcc
     2221 gcacataccg cctgagtctc ctggaggagt acggctgctg caaggagctg gccagccggc
     2281 tccgctacgc ccgcactatg gtggacaagc tgctgagctc acgctcggcc agcaaccgtc
     2341 tcaaggcctc cggtagcggc agcgcatccg gcggaagcgg aagcggaccg gtcgccacca
     2401 tggtgagtgt gattaaacca gagatgaaga tgaggtacta catggacggc tccgtcaatg
     2461 ggcatgagtt cacaattgaa ggtgaaggca caggcagacc ttacgaggga catcaagaga
     2521 tgacactacg cgtcacaatg gccgagggcg ggccaatgcc tttcgcgttt gacttagtgt
     2581 cacacgtgtt ctgttacggc cacagagtat ttactaaata tccagaagag ataccagact
     2641 atttcaaaca agcatttcct gaaggcctgt catgggaaag gtcgttggag ttcgaagatg
     2701 gtgggtccgc ttcagtcagt gcgcatataa gccttagagg aaacaccttc taccacaaat
     2761 ccaaatttac tggggttaac tttcctgccg atggtcctat catgcaaaac caaagtgttg
     2821 attgggagcc atcaaccgag aaaattactg ccagcgacgg agttctgaag ggtgatgtta
     2881 cgatgtacct aaaacttgaa ggaggcggca atcacaaatg ccaaatgaag actacttaca
     2941 aggcggcaaa agagattctt gaaatgccag gagaccatta catcggccat cgcctcgtca
     3001 ggaaaaccga aggcaacatt actgagcagg tagaagatgc agtagctcat agctaagcgg
     3061 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3121 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3181 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3241 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3301 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3361 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3421 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3481 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3541 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3601 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3661 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3721 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3781 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3841 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3901 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3961 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4021 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4081 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4141 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4201 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4261 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4321 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4381 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4441 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4501 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4561 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4621 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4681 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4741 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4801 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4861 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4921 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4981 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5041 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5101 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5161 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5221 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5281 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5341 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5401 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5461 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5521 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5581 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5641 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5701 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5761 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5821 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5881 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5941 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6001 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6061 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6121 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6181 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6241 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6301 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6361 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     6421 attagttcat agcccatata tggagttccg
//
