LOCUS       sequence-162751-        4691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: N1 Cloning Vector, Excitation: 508 / 572, Emission: 
            518 / 580.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4691)
  AUTHORS   Kremers GJ, Hazelwood KL, Murphy CS, Davidson MW, Piston DW
  TITLE     Photoconversion in orange and red fluorescent proteins.
  JOURNAL   Nat Methods. 2009 May;6(5):355-8. doi: 10.1038/nmeth.1319. Epub 2009
            Apr 12.
  PUBMED    19363494
REFERENCE   2  (bases 1 to 4691)
  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"; doi: 
            "10.1038/nmeth.1319"; journalName: "Nat Methods"; date: "2009-05"; 
            volume: "6"; issue: "5"; pages: "355-8"
COMMENT     Sequence Label: Kaede-N1
FEATURES             Location/Qualifiers
     source          1..4691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    546..626
                     /label=MCS
                     /note="multiple cloning site"
     CDS             634..1311
                     /codon_start=1
                     /product="Kaede green-to-red photoconvertible fluorescent 
                     protein (Ando et al., 2002)"
                     /label=Kaede
                     /translation="MSLIKPEMKIKLLMEGNVNGHQFVIEGDGKGHPFEGKQSMDLVVK
                     EGAPLPFAYDILTTAFHYGNRVFAKYPDHIPDYFKQSFPKGFSWERSLMFEDGGVCIAT
                     NDITLKGDTFFNKVRFDGVNFPPNGPVMQKKTLKWEASTEKMYLRDGVLTGDITMALLL
                     KGDVHYRCDFRTTYKSRQEGVKLPGYHFVDHCISILRHDKDYNEVKLYEHAVAHSGLPD
                     NVK"
     polyA_signal    1434..1555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1471..1490)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1525..1544
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1562..2017)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1699..1720)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1911..1930
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2044..2148
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2150..2507
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2150..2341
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2173..2193)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2358..2493
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2542..3336
                     /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             2542..3336
                     /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(2596..2615)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3206..3225
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3524..3543)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3568..3615
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3944..4532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4433..4452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgacg
      601 gtaccgcggg cccgggatcc accggtcgcc accatgagtc tgattaaacc agaaatgaag
      661 atcaagctgc ttatggaagg caatgtaaac gggcaccagt ttgttattga gggagatgga
      721 aaaggccatc cttttgaggg aaaacagagt atggaccttg tagtcaaaga aggcgcacct
      781 ctcccttttg cctacgatat cttgacaaca gcattccatt atggtaacag ggtttttgct
      841 aaatacccag accatatacc agactacttc aagcagtcgt ttcccaaagg gttttcttgg
      901 gagcgaagcc tgatgttcga ggacgggggc gtttgcatcg ctacaaatga cataacactg
      961 aaaggagaca ctttttttaa caaagttcga tttgatggcg taaactttcc cccaaatggt
     1021 cctgttatgc agaagaagac tctgaaatgg gaggcatcca ctgagaaaat gtatttgcgt
     1081 gatggagtgt tgacgggcga tattacaatg gctctgctgc ttaaaggaga tgtccattac
     1141 cgatgtgact tcagaactac ttacaaatct aggcaggagg gtgtcaagtt gccaggatat
     1201 cactttgtcg atcactgcat cagcatattg aggcatgaca aagactacaa cgaggttaag
     1261 ctgtatgagc acgctgttgc ccattctgga ttgccggaca acgtcaagta aagcggccgc
     1321 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1381 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1441 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1501 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     1561 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     1621 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1681 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1741 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     1801 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     1861 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     1921 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     1981 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2041 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2101 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     2161 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2221 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2281 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2341 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2401 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2461 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     2521 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     2581 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     2641 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     2701 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     2761 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     2821 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     2881 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     2941 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3001 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3061 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3121 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3181 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3241 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3301 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     3361 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     3421 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     3481 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     3541 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     3601 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     3661 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     3721 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     3781 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3841 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3901 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3961 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4021 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4081 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4141 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4201 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4261 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4321 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4381 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4441 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4501 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4561 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4621 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta
     4681 gttcatagcc c
//
