LOCUS       sequence-232476-        4779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Lysosomes/ER/Golgi, Excitation: 488, Emission: 507.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4779)
  TITLE     Davidson Green fluorescent proteins
REFERENCE   2  (bases 1 to 4779)
  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: EGFP-CathespinB-6
FEATURES             Location/Qualifiers
     source          1..4779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        159..350
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     307..327
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(352..456)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      483..938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(570..589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     780..801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    945..1066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(956..975)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1010..1029
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1248..1958,1959..1961,1962..1964))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1290..1311)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1639..1658
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1898..1919
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1961..1970
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2065..2268)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2094..2114)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2269..2572
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2747..3335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2827..2846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3664..3711
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3736..3755
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3943..4737)
                     /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             complement(3943..4737)
                     /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(4054..4073)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4664..4683
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4772..4779,1..350))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
       61 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
      121 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
      181 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
      241 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
      301 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag gactcttcct
      361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
      421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
      481 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
      541 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      601 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
      661 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
      721 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
      781 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
      841 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
      901 taacgcgaat tttaacaaaa tattaacgct tacaatttac gcgttaagat acattgatga
      961 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1021 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1081 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1141 cctctacaaa tgtggtatgg ctgattatga tcagttatct agatccggtg gatcccgggc
     1201 ccgcggtacc gtcgactgca gaattcttac agctcgtcct ttccggactt gtacagctcg
     1261 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     1321 ttctcgttgg ggtctttgct cagcttggac tgggtgctca ggtagtggtt gtcgggcagc
     1381 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg
     1441 ccgtcctcga tgttgcagtt gaggccgaag ttggccttga tgccgttctt ctgcttgtcg
     1501 gcggtgatat agacgttgcc gtggatggcg ttgtactcca gcttgtgccc caggatgttg
     1561 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     1621 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     1681 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     1741 tagcgggcga agcactgcac gccccagctc agggtggtca cgagggtggg ccagggcacg
     1801 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc
     1861 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     1921 atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc gaccggtgcg
     1981 gcgacggcca ggccgaggag gccgagcagc aacggcacgg atagcagcat ggtggcgcta
     2041 gcggatctga cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct
     2101 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
     2161 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     2221 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
     2281 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     2341 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     2401 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     2461 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     2521 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     2581 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac taatgcatgg
     2641 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2701 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2761 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2821 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2881 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2941 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3001 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3061 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3121 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3181 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3241 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3301 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3361 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3421 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3481 tatatgagta acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg
     3541 ggccttcacc cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc
     3601 cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt
     3661 tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc
     3721 gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga
     3781 agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga
     3841 ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt
     3901 tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag
     3961 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     4021 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     4081 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     4141 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     4201 gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     4261 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     4321 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     4381 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     4441 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     4501 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag
     4561 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     4621 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     4681 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     4741 aaacgatcct catcctgtct cttgatcgat ctttgcaaa
//
