LOCUS       sequence-226429-        5700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Lysosomes, Excitation: 549, Emission: 565.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5700)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 5700)
  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: mOrange2-CathespinB-6
FEATURES             Location/Qualifiers
     source          1..5700
                     /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      1590..1599
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1596..2306
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    2428..2549
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2465..2484)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2519..2538
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2556..3011)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2693..2714)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2905..2924
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3038..3142
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3144..3501
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3144..3335
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3167..3187)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3352..3487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3536..4330
                     /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             3536..4330
                     /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(3590..3609)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4200..4219
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4518..4537)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4562..4609
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4938..5526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5427..5446
                     /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 atctcgagct caagcttcga attccaggat gtggtggtcc ttgatccctc
      601 tctcttgcct gctggcactg accagtgccc atgacaagcc ttcctttcac ccactgtcgg
      661 acgacatgat taactatatc aacaaacaga atacaacatg gcaggctgga cgcaacttct
      721 acaatgttga cataagctat ctgaagaagc tgtgtggaac tgtcctgggt ggacccaagc
      781 tgccggaaag ggttgggttc agcgaggaca taaatctacc tgaatccttt gatgcacggg
      841 aacagtggtc caattgcccg accatcgcac agatcagaga ccaggggtcc tgtggctctt
      901 gttgggcatt tggggcagtg gaagccatgt ctgaccgaat ctgcattcac accaatggcc
      961 gagtcaatgt ggaggtgtct gctgaggacc tgcttacctg ctgtggtatc cagtgtgggg
     1021 atggctgtaa tggtggctat ccctctggag catggaactt ctggactaga aaaggcctgg
     1081 tttctggtgg agtatacaat tctcatatag gctgcttacc ctacaccatc cctccctgtg
     1141 aacaccatgt caatggctcc cgtcccccat gcactggaga aggagatact cccaagtgca
     1201 acaagatgtg tgaggctggc tactccacat cctacaagga agataagcac tatgggtaca
     1261 cttcctacag tgtgtctgac agcgagaagg agatcatggc ggaaatctac aaaaatggcc
     1321 cagtggaggg tgcttttact gtgttttctg acttcttgac ttacaaatca ggcgtataca
     1381 agcatgaagc cggtgatgtg atgggaggcc atgccatccg cattctgggc tggggaatag
     1441 agaatggagt accctactgg ctggtagcaa actcctggaa cgttgactgg ggtgataatg
     1501 gtttctttaa aatcctcaga ggagagaacc actgtggaat tgaatcagaa atcgtggctg
     1561 gaatcccacg cactcaggat ccaccggtcg ccaccatggt gagcaagggc gaggagaata
     1621 acatggccat catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg
     1681 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc tttcagaccg
     1741 ctaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     1801 atttcaccta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttca
     1861 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaactacgag gacggcggcg
     1921 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     1981 tgcgcggcac caacttcccc tccgacggcc ccgtgatgca gaagaagacc atgggctggg
     2041 aggcctcctc cgagcggatg taccccgagg acggtgccct gaagggcaag atcaagatga
     2101 ggctgaagct gaaggacggc ggccactaca cctccgaggt caagaccacc tacaaggcca
     2161 agaagcccgt gcagctgccc ggcgcctaca tcgtcgacat caagttggac atcacctccc
     2221 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     2281 gcggcatgga cgagctgtac aagtaagcgg ccgcgactct agatcataat cagccatacc
     2341 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2401 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2461 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2521 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     2581 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2641 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2701 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2761 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2821 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2881 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2941 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3001 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3061 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3121 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3181 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3241 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3301 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3361 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3421 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3481 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3541 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3601 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3661 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3721 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3781 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3841 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3901 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3961 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4021 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4081 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4141 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4201 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4261 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4321 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4381 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4441 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4501 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4561 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     4621 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4681 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4741 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     4801 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4861 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4921 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4981 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5041 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5101 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5161 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5221 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5281 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5341 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5401 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5461 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5521 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5581 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     5641 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
