LOCUS       sequence-116536-        5817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Golgi-CNL in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5817)
  AUTHORS   Takai A, Nakano M, Saito K, Haruno R, Watanabe TM, Ohyanagi T, Jin 
            T, Okada Y, Nagai T
  TITLE     Expanded palette of Nano-lanterns for real-time multicolor 
            luminescence imaging.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):4352-6. doi: 
            10.1073/pnas.1418468112. Epub 2015 Mar 23.
  PUBMED    25831507
REFERENCE   2  (bases 1 to 5817)
  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: 
            "10.1073/pnas.1418468112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-04-7- 7"; volume: "112"; issue: "14"; pages: "4352-6"
COMMENT     Sequence Label: pCNL-N1_Golgi
FEATURES             Location/Qualifiers
     source          1..5817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      858..867
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(864..866,867..869,870..1382)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(909..930)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1170..1189)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1517..1538
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2605..2726
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2642..2661)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2696..2715
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2733..3188)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2870..2891)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3082..3101
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3215..3319
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3321..3678
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3321..3512
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3344..3364)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3529..3664
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3713..4507
                     /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             3713..4507
                     /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(3767..3786)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4377..4396
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4695..4714)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4739..4786
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5115..5703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5604..5623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatga
      601 ggcttcggga gccgctcctg agcggcagcg ccgcgatgcc aggcgcgtcc ctacagcggg
      661 cctgccgcct gctcgtggcc gtctgcgctc tgcaccttgg cgtcaccctc gtttactacc
      721 tggctggccg cgacctgagc cgcctgcccc aactggtcgg agtctccaca ccgctgcagg
      781 gcggctcgaa cagtgccgcc gccatcgggc agtcctccgg ggagctccgg accggagggg
      841 ccaaggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      901 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      961 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1021 agctgcccgt gccctggccc accctcgtga ccaccctgtc ctggggcgtg cagtgcttcg
     1081 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1141 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1201 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1261 aggacggcaa catcctgggg cacaagctgg agtacaacta ctttagcgac aacgtctata
     1321 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     1381 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1441 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     1501 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg ggtaccaagg
     1561 tgtacgaccc cgagcagagg aagaggatga tcaccggccc ccagtggtgg gccaggtgca
     1621 agcagatgaa cgtgctggac agcttcatca actactacga cagcgagaag cacgccgaga
     1681 acgccgtgat cttcctgcac ggcaacgcca ctagcagcta cctgtggagg cacgtggtgc
     1741 cccacatcga gcccgtggcc aggtgcatca tccccgatct gatcggcatg ggcaagagcg
     1801 gcaagagcgg caacggcagc tacaggctgc tggaccacta caagtacctg accgcctggt
     1861 tcgagcttct gaacctgccc aagaagatca tcttcgtggg ccacgactgg ggcgccgccc
     1921 tggccttcca ctacgcctac gagcaccagg acaggatcaa ggccatcgtg cacatggaga
     1981 gcgtggtgga cgtgatcgag agctgggacg agtggccaga catcgaggag gacatcgccc
     2041 tgatcaagag cgaggagggc gagaagatgg tgctggagaa caacttcttc gtggagaccg
     2101 tgctgcccag caagatcatg agaaagctgg agcccgagga gttcgccgcc tacctggagc
     2161 ccttcaagga gaagggcgag gtgagaagac ccaccctgag ctggcccaga gagatccccc
     2221 tggtgaaggg cggcaagccc gacgtggtgc agatcgtgag aaactacaac gcctacctga
     2281 gagccagcga cgacctgccc aagctgttca tcgagggcga ccccggcttc ttcagcaacg
     2341 ccatcgtgga gggcgccaag aagttcccca acaccgagtt cgtgaaggtg aagggcctgc
     2401 acttcctcca ggaggacgcc cccgacgaga tgggcaagta catcaagagc ttcgtggaga
     2461 gagtgctgaa gaacgagcag taggcggccg cgactctaga tcataatcag ccataccaca
     2521 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     2581 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     2641 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     2701 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     2761 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     2821 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     2881 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     2941 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3001 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3061 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3121 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3181 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3241 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3301 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3361 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3421 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3481 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3541 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3601 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3661 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     3721 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3781 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3841 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     3901 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3961 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4021 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4081 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4141 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4201 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4261 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4321 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4381 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4441 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4501 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4561 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4621 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     4681 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4741 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     4801 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4861 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     4921 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     4981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5221 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5761 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
