LOCUS       sequence-241031-        7004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Transferrin Receptor, Excitation: 548, Emission: 561.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7004)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 7004)
  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-TFR-20
FEATURES             Location/Qualifiers
     source          1..7004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        41..344
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        345..548
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     499..519
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2940..2949
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2973..3623
                     /codon_start=1
                     /product="monomeric Kusabira-Orange fluorescent protein 
                     (Karasawa et al., 2004)"
                     /label=mKO
                     /translation="MAVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     polyA_signal    3772..3893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3809..3828)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3863..3882
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3900..4355)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4037..4058)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4249..4268
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4382..4486
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4488..4845
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4488..4679
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4511..4531)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4696..4831
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4880..5674
                     /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             4880..5674
                     /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(4934..4953)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5544..5563
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5862..5881)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5906..5953
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6282..6870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6771..6790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
       61 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      121 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      181 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      241 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      301 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      361 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      421 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      481 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      541 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagcg
      601 ccaccatgat ggatcaagct agatcagcat tctctaactt gtttggtgga gaaccattgt
      661 catatacccg gttcagcctg gctcggcaag tagatggcga taacagtcat gtggagatga
      721 aacttgctgt agatgaagaa gaaaatgctg acaataacac aaaggccaat gtcacaaaac
      781 caaaaaggtg tagtggaagt atctgctatg ggactattgc tgtgatcgtc tttttcttga
      841 ttggatttat gattggctac ttgggctatt gtaaaggggt agaaccaaaa actgagtgtg
      901 agagactggc aggaaccgag tctccagtga gggaggagcc aggagaggac ttccctgcag
      961 cacgtcgctt atattgggat gacctgaaga gaaagttgtc ggagaaactg gacagcacag
     1021 acttcaccag caccatcaag ctgctgaatg aaaattcata tgtccctcgt gaggctggat
     1081 ctcaaaaaga tgaaaatctt gcgttgtatg ttgaaaatca atttcgtgaa tttaaactca
     1141 gcaaagtctg gcgtgatcaa cattttgtta agattcaggt caaagacagc gctcaaaact
     1201 cggtgatcat agttgataag aacggtagac ttgtttacct ggtggagaat cctgggggtt
     1261 atgtggcgta tagtaaggct gcaacagtta ctggtaaact ggtccatgct aattttggta
     1321 ctaaaaaaga ttttgaggat ttatacactc ctgtgaatgg atctatagtg attgtcagag
     1381 cagggaaaat cacctttgca gaaaaggttg caaatgctga aagcttaaat gcaattggtg
     1441 tgttgatata catggaccag actaaatttc ccattgttaa cgcagaactt tcattctttg
     1501 gacatgctca tctggggaca ggtgaccctt acacacctgg attcccttcc ttcaaccaca
     1561 ctcagtttcc accatctcgg tcatcaggat tgcctaatat acctgtccag acaatctcca
     1621 gagctgctgc agaaaagctg tttgggaata tggaaggaga ctgtccctct gactggaaaa
     1681 cagactctac atgtaggatg gtaacctcag aaagcaagaa tgtgaagctc actgtgagca
     1741 atgtgctgaa agagataaaa attcttaaca tctttggagt tattaaaggc tttgtagaac
     1801 cagatcacta tgttgtagtt ggggcccaga gagatgcatg gggccctgga gctgcaaaat
     1861 ccggtgtagg cacagctctc ctattgaaac ttgcccagat gttctcagat atggtcttaa
     1921 aagatgggtt tcagcccagc agaagcatta tctttgccag ttggagtgct ggagactttg
     1981 gatcggttgg tgccactgaa tggctagagg gatacctttc gtccctgcat ttaaaggctt
     2041 tcacttatat taatctggat aaagcggttc ttggtaccag caacttcaag gtttctgcca
     2101 gcccactgtt gtatacgctt attgagaaaa caatgcaaaa tgtgaagcat ccggttactg
     2161 ggcaatttct atatcaggac agcaactggg ccagcaaagt tgagaaactc actttagaca
     2221 atgctgcttt ccctttcctt gcatattctg gaatcccagc agtttctttc tgtttttgcg
     2281 aggacacaga ttatccttat ttgggtacca ccatggacac ctataaggaa ctgattgaga
     2341 ggattcctga gttgaacaaa gtggcacgag cagctgcaga ggtcgctggt cagttcgtga
     2401 ttaaactaac ccatgatgtt gaattgaacc tggactatga gaggtacaac agccaactgc
     2461 tttcatttgt gagggatctg aaccaataca gagcagacat aaaggaaatg ggcctgagtt
     2521 tacagtggct gtattctgct cgtggagact tcttccgtgc tacttccaga ctaacaacag
     2581 atttcgggaa tgctgagaaa acagacagat ttgtcatgaa gaaactcaat gatcgtgtca
     2641 tgagagtgga gtatcacttc ctctctccct acgtatctcc aaaagagtct cctttccgac
     2701 atgtcttctg gggctccggc tctcacacgc tgccagcttt actggagaac ttgaaactgc
     2761 gtaaacaaaa taacggtgct tttaatgaaa cgctgttcag aaaccagttg gctctagcta
     2821 cttggactat tcagggagct gcaaatgccc tctctggtga cgtttgggac attgacaatg
     2881 agttttcaga attcggctcc accggctcca ccggctccac cggcgcggat ccaccggtcg
     2941 ccaccatggt gagcaagggc gaggagaata acatggccgt gattaaacca gagatgaaga
     3001 tgaggtacta catggacggc tccgtcaatg ggcatgagtt cacaattgaa ggtgaaggca
     3061 caggcagacc ttacgaggga catcaagaga tgacactacg cgtcacaatg gccaagggcg
     3121 ggccaatgcc tttcgcgttt gacttagtgt cacacgtgtt ctgttacggc cacagacctt
     3181 ttactaaata tccagaagag ataccagact atttcaaaca agcatttcct gaaggcctgt
     3241 catgggaaag gtcgttggag ttcgaagatg gtgggtccgc ttcagtcagt gcgcatataa
     3301 gccttagagg aaacaccttc taccacaaat ccaaatttac tggggttaac tttcctgccg
     3361 atggtcctat catgcaaaac caaagtgttg attgggagcc atcaaccgag aaaattactg
     3421 ccagcgacgg agttctgaag ggtgatgtta cgatgtacct aaaacttgaa ggaggcggca
     3481 atcacaaatg ccaattcaag actacttaca aggcggcaaa aaagattctt aaaatgccag
     3541 gaagccatta catcagccat cgcctcgtca ggaaaaccga aggcaacatt actgagctgg
     3601 tagaagatgc agtagctcat tccggcggca tggacgagct gtacaagtaa gcggccgcga
     3661 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3721 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3781 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3841 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     3901 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3961 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     4021 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     4081 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     4141 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     4201 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     4261 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4321 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     4381 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4441 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     4501 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4561 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4621 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4681 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4741 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4801 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4861 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4921 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4981 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5041 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5101 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5161 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5221 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5281 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5341 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5401 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5461 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5521 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5581 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5641 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5701 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5761 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5821 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     5881 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5941 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     6001 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     6061 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     6121 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6181 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6241 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6301 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6361 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6421 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6481 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6541 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6601 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6661 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6721 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6781 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6841 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6901 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6961 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaa
//
