LOCUS       Exported                4722 bp ds-DNA     circular SYN 09-SEP-2021
DEFINITION  Localization: C1 Cloning Vector, Excitation: 568, Emission: 592.
ACCESSION   .
VERSION     .
KEYWORDS    mApple-C1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4722)
  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 4722)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 9, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4722
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(5..25)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      190..325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     252..271
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             374..1168
                     /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(428..447)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1038..1057
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1356..1375)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1400..1447
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1776..2364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2265..2284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2539..2842
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2843..3046
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2997..3017
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3085..3094
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3091..3798
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     3504..3523
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3799..3864
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3988..4109
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4025..4044)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4079..4098
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4116..4571)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4253..4274)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4465..4484
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4598..4702
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4704..339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
       61 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
      121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
      181 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
      241 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
      301 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
      361 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
      421 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
      481 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
      541 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
      601 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
      661 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
      721 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
      781 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
      841 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
      901 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
      961 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     1021 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     1081 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     1141 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     1201 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     1261 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     1321 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     1381 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     1441 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     1501 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     1561 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     1621 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1681 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1741 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1801 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1861 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1921 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1981 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2041 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2101 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2161 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2221 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2281 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2341 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2401 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2461 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     2521 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2581 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2641 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2701 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2761 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2821 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     2881 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     2941 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     3001 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     3061 tcagatccgc tagcgctacc ggtcgccacc atggtgagca agggcgagga gaataacatg
     3121 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     3181 gagttcgaga tcgagggcga gggcgagggc cgcccctacg aggcctttca gaccgctaag
     3241 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     3301 atgtacggct ccaaggtcta cattaagcac ccagccgaca tccccgacta cttcaagctg
     3361 tccttccccg agggcttcag gtgggagcgc gtgatgaact tcgaggacgg cggcattatt
     3421 cacgttaacc aggactcctc cctgcaggac ggcgtgttca tctacaaggt gaagctgcgc
     3481 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     3541 tccgaggagc ggatgtaccc cgaggacggc gccctgaaga gcgagatcaa gaagaggctg
     3601 aagctgaagg acggcggcca ctacgccgcc gaggtcaaga ccacctacaa ggccaagaag
     3661 cccgtgcagc tgcccggcgc ctacatcgtc gacatcaagt tggacatcgt gtcccacaac
     3721 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     3781 atggacgagc tgtacaagtc cggactcaga tctcgagctc aagcttcgaa ttctgcagtc
     3841 gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc
     3901 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3961 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4021 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4081 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     4141 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4201 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4261 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4321 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4381 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4441 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4501 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4561 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4621 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4681 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc ag
//
