LOCUS       601-pCre-P2A-mCherry        5864 bp    DNA     circular SYN  7-APR-2023
DEFINITION  Encodes P2A-mCherry preceded by a multiple cloning sequence. Enables bicistronic expression of protein of interest and mCherry.
SOURCE      synthetic DNA construct.
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4790)
  AUTHORS   van Unen J, Rashidfarrokhi A, Hoogendoorn E, Postma M, Gadella TW 
            Jr, Goedhart J
  TITLE     Quantitative Single-Cell Analysis of Signaling Pathways Activated 
            Immediately Downstream of Histamine Receptor Subtypes.
  JOURNAL   Mol Pharmacol. 2016 Sep;90(3):162-76. doi: 10.1124/mol.116.104505. 
            Epub 2016 Jun 29.
REFERENCE   2  (bases 1 to 4790)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 7, 2023 from SnapGene Viewer 6.2.1 to Vector NTI(R) 
            format
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1124/mol.116.104505"; journalName: "Mol Pharmacol"; date: 
            "2016-09"; volume: "90"; issue: "3"; pages: "162-76"
COMMENT     This file is created by Vector NTI
            http://www.invitrogen.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTDATE|-14864805|
COMMENT     VNTDBDATE|-14864807|
COMMENT     LSOWNER|
COMMENT     VNTNAME|601-pCre-P2A-mCherry|
COMMENT     VNTAUTHORNAME|Daekee|
COMMENT     VNTAUTHORTEL|822-3277-4368|
COMMENT     VNTAUTHOREML|daekee@ewha.ac.kr|
COMMENT     VNTOAUTHORNAME|UNKNOWN|
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COMMENT                          "DNA '601-pCre-P2A-mCherry'" 1) 
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COMMENT                          "Encodes P2A-mCherry preceded by a multiple cloning sequence. Enables bicistronic expression of protein of interest and mCherry." 
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COMMENT                          "Currently local object. Original author: UNKNOWN" 1) 
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COMMENT                          "Created: 04/07/23 05:00\277\300\310\304" 1) 
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COMMENT                          "  AUTHORS   van Unen J, Rashidfarrokhi A, Hoogendoorn E, Postma M, Gadella TW " 
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COMMENT                          "  TITLE     Quantitative Single-Cell Analysis of Signaling Pathways Activated " 
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COMMENT                          "            Immediately Downstream of Histamine Receptor Subtypes." 
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COMMENT                          "  JOURNAL   Mol Pharmacol. 2016 Sep;90(3):162-76. doi: 10.1124/mol.116.104505. " 
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COMMENT                          "            Epub 2016 Jun 29." 1) 
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FEATURES             Location/Qualifiers
     enhancer        61..364
                     /vntifkey="9"
                     /label=CMV\enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /vntifkey="30"
                     /label=CMV\promoter
                     /note="human cytomegalovirus (CMV) immediate early promoter"
     CDS             1756..1812
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /vntifkey="4"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides."
     CDS             1819..2529
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /vntifkey="4"
                     /label=mCherry
                     /note="mammalian codon-optimized"
     polyA_signal    2652..2773
                     /vntifkey="25"
                     /label=SV40\poly(A)\signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2780..3235)
                     /direction=LEFT
                     /vntifkey="33"
                     /label=f1\ori
                     /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
     promoter        3368..3725
                     /vntifkey="30"
                     /label=SV40\promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3576..3711
                     /vntifkey="33"
                     /label=SV40\ori
                     /note="SV40 origin of replication"
     CDS             3760..4554
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /vntifkey="4"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))"
     polyA_signal    4786..4833
                     /vntifkey="25"
                     /label=HSV\TK\poly(A)\signal
                     /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5162..5750
                     /direction=RIGHT
                     /vntifkey="33"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     CDS             670..1698
                     /vntifkey="4"
                     /label=Cre
     misc_feature    670..672
                     /vntifkey="21"
                     /label=START
BASE COUNT     1418 a      1528 c      1583 g      1335 t 
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 gctagcgcta 
      601 ccggactcag atctcgaggg gcagagccga tcctgtacac tttacttaaa accattatct 
      661 gagtgtgaaa tgtccaattt actgaccgta caccaaaatt tgcctgcatt accggtcgat 
      721 gcaacgagtg atgaggttcg caagaacctg atggacatgt tcagggatcg ccaggcgttt 
      781 tctgagcata cctggaaaat gcttctgtcc gtttgccggt cgtgggcggc atggtgcaag 
      841 ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg ttcgcgatta tcttctatat 
      901 cttcaggcgc gcggtctggc agtaaaaact atccagcaac atttgggcca gctaaacatg 
      961 cttcatcgtc ggtccgggct gccacgacca agtgacagca atgctgtttc actggttatg 
     1021 cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgtg caaaacaggc tctagcgttc 
     1081 gaacgcactg atttcgacca ggttcgttca ctcatggaaa atagcgatcg ctgccaggat 
     1141 atacgtaatc tggcatttct ggggattgct tataacaccc tgttacgtat agccgaaatt 
     1201 gccaggatca gggttaaaga tatctcacgt actgacggtg ggagaatgtt aatccatatt 
     1261 ggcagaacga aaacgctggt tagcaccgca ggtgtagaga aggcacttag cctgggggta 
     1321 actaaactgg tcgagcgatg gatttccgtc tctggtgtag ctgatgatcc gaataactac 
     1381 ctgttttgcc gggtcagaaa aaatggtgtt gccgcgccat ctgccaccag ccagctatca 
     1441 actcgcgccc tggaagggat ttttgaagca actcatcgat tgatttacgg cgctaaggat 
     1501 gactctggtc agagatacct ggcctggtct ggacacagtg cccgtgtcgg agccgcgcga 
     1561 gatatggccc gcgctggagt ttcaataccg gagatcatgc aagctggtgg ctggaccaat 
     1621 gtaaatattg tcatgaacta tatccgtaac ctggatagtg aaacaggggc aatggtgcgc 
     1681 ctgctggaag atggcgatgt cgaaattctg cagtcgacgg taccgcgggc ccgggatcca 
     1741 ccggtgggat cgggtgctac taacttcagc ctgctgaagc aggctggaga cgtggaggag 
     1801 aaccctggac ctgggtccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag 
     1861 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc 
     1921 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc 
     1981 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc 
     2041 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag 
     2101 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag 
     2161 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc 
     2221 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg 
     2281 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac 
     2341 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg 
     2401 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc 
     2461 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg 
     2521 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt 
     2581 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa 
     2641 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 
     2701 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 
     2761 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat 
     2821 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa 
     2881 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta 
     2941 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca 
     3001 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat 
     3061 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg 
     3121 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc 
     3181 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt 
     3241 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 
     3301 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 
     3361 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt 
     3421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 
     3481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 
     3541 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 
     3601 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 
     3661 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 
     3721 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 
     3781 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 
     3841 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 
     3901 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta 
     3961 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 
     4021 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 
     4081 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 
     4141 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 
     4201 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 
     4261 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc 
     4321 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 
     4381 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 
     4441 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 
     4501 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 
     4561 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 
     4621 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 
     4681 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa 
     4741 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac 
     4801 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct 
     4861 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct 
     4921 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg 
     4981 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc 
     5041 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 
     5101 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 
     5161 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 
     5221 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 
     5281 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact 
     5341 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 
     5401 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 
     5461 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 
     5521 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 
     5581 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 
     5641 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 
     5701 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 
     5761 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 
     5821 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat 
//
