LOCUS       sequence-115869-        5636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CNL-PTS1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5636)
  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 5636)
  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-C1_PTS1
FEATURES             Location/Qualifiers
     source          1..5636
                     /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      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(603..605,606..608,609..1121)
                     /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(648..669)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(909..928)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1256..1277
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2424..2545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2461..2480)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2515..2534
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2552..3007)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2689..2710)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2901..2920
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3034..3138
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3140..3497
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3140..3331
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3163..3183)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3348..3483
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3532..4326
                     /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             3532..4326
                     /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(3586..3605)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4196..4215
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4514..4533)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4558..4605
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4934..5522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5423..5442
                     /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 gctagcgcca
      601 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      661 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      721 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      781 ccctcgtgac caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga
      841 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      901 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      961 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1021 acaagctgga gtacaactac tttagcgaca acgtctatat caccgccgac aagcagaaga
     1081 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     1141 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1201 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     1261 tcctgctgga gttcgtgacc gccgccgggg gtaccaaggt gtacgacccc gagcagagga
     1321 agaggatgat caccggcccc cagtggtggg ccaggtgcaa gcagatgaac gtgctggaca
     1381 gcttcatcaa ctactacgac agcgagaagc acgccgagaa cgccgtgatc ttcctgcacg
     1441 gcaacgccac tagcagctac ctgtggaggc acgtggtgcc ccacatcgag cccgtggcca
     1501 ggtgcatcat ccccgatctg atcggcatgg gcaagagcgg caagagcggc aacggcagct
     1561 acaggctgct ggaccactac aagtacctga ccgcctggtt cgagcttctg aacctgccca
     1621 agaagatcat cttcgtgggc cacgactggg gcgccgccct ggccttccac tacgcctacg
     1681 agcaccagga caggatcaag gccatcgtgc acatggagag cgtggtggac gtgatcgaga
     1741 gctgggacga gtggccagac atcgaggagg acatcgccct gatcaagagc gaggagggcg
     1801 agaagatggt gctggagaac aacttcttcg tggagaccgt gctgcccagc aagatcatga
     1861 gaaagctgga gcccgaggag ttcgccgcct acctggagcc cttcaaggag aagggcgagg
     1921 tgagaagacc caccctgagc tggcccagag agatccccct ggtgaagggc ggcaagcccg
     1981 acgtggtgca gatcgtgaga aactacaacg cctacctgag agccagcgac gacctgccca
     2041 agctgttcat cgagggcgac cccggcttct tcagcaacgc catcgtggag ggcgccaaga
     2101 agttccccaa caccgagttc gtgaaggtga agggcctgca cttcctccag gaggacgccc
     2161 ccgacgagat gggcaagtac atcaagagct tcgtggagag agtgctgaag aacgagcagt
     2221 ccggactcag atctcgagct caagcttcga attccgggat atccaagctg tgagtcgacg
     2281 gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc cataccacat
     2341 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     2401 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2461 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2521 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     2581 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2641 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2701 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2761 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2821 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2881 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     2941 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3001 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3061 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3121 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3181 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3241 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3301 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3361 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3421 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3481 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3541 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3601 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3661 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     3721 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3781 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3841 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3901 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3961 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4021 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4081 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4141 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4201 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4261 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4321 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4381 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4441 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4501 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4561 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4621 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4681 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4741 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4801 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4861 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4921 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4981 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5041 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5101 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5161 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5221 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5281 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5341 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5401 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5461 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5521 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5581 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
