LOCUS       Exported                7803 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    CMV-Brainbow-1.0 H
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7803)
  AUTHORS   Livet J, Weissman TA, Kang H, Draft RW, Lu J, Bennis RA, Sanes JR, 
            Lichtman JW
  TITLE     Transgenic strategies for combinatorial expression of fluorescent 
            proteins in the nervous system.
  JOURNAL   Nature. 2007 Nov 1. 450(7166):56-62.
  PUBMED    17972876
REFERENCE   2  (bases 1 to 7803)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7803
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    657..690
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    707..740
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      757..766
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      761..770
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(2214..2234)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2214..2232)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2252..2269)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2258..2369
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    2528..2561
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     regulatory      2680..2689
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2686..3405
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3528..3649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3664..3697
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      3820..3829
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3826..4545
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    4588..4621
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    4645..4766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4773..5228)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4910..4931)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5122..5141
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5255..5359
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5361..5718
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(5384..5404)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5569..5704
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5631..5650
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5753..6547
                     /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(5807..5826)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6417..6436
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6735..6754)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6779..6826
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7155..7743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7644..7663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gagctcataa
      661 cttcgtatag gatactttat acgaagttat aacgttagat ctcgagataa cttcgtatag
      721 catacattat acgaagttat accggctagt gatttcgcca ccatggtggc ctcctccgag
      781 gacgtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac
      841 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
      901 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc
      961 cagtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgacta caagaagctg
     1021 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     1081 accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagttccgc
     1141 ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     1201 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg
     1261 aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag
     1321 cccgtgcagc tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac
     1381 gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg
     1441 gggcatggca ccggcagcac cggcagcggc agctccggca ccgcctcctc cgaggacgtc
     1501 atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg ccacgagttc
     1561 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     1621 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttccagtac
     1681 ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg actacaagaa gctgtccttc
     1741 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     1801 acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagtt ccgcggcacc
     1861 aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc
     1921 gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg
     1981 aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg
     2041 cagctgcccg gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac
     2101 tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt cctgtaggaa
     2161 ttggccgcga ctctaggatt gatctagagt cgcggctaga taatagggta ccacctttag
     2221 tgagggttaa ttgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     2281 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tctcactgtc
     2341 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2401 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2461 ggggcgccgc tccgagggcc gccaccacct gttcctgtag gaattggccg cgactctagg
     2521 atctaggata acttcgtata ggatacttta tacgaagtta ttgatcaatt cacagccacc
     2581 atgctgtgct gcatcagaag aactaaaccg gttgagaaga atgaagaggc cgatcaggag
     2641 ctgcagtcga cggtaccgcg ggcccgggat ccaccggtcg ccaccatggt gagcaagggc
     2701 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2761 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2821 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
     2881 ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2941 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     3001 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     3061 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3121 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     3181 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     3241 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     3301 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3361 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct
     3421 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3481 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3541 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3601 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgcgattc
     3661 gagataactt cgtatagcat acattatacg aagttatacc ggaattaatt cacagccacc
     3721 atgctgtgct gcatcagaag aactaaaccg gttgagaaga atgaagaggc cgatcaggag
     3781 ctgcagtcga cggtaccgcg ggcccgggat ccaccggtcg ccaccatggt gagcaagggc
     3841 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     3901 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     3961 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     4021 acctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     4081 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     4141 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     4201 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     4261 gccatcagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     4321 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     4381 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     4441 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     4501 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct
     4561 agatcaccgc gactctagat cacgcgcgaa gttcctattc tctagaaagt ataggaactt
     4621 ctcgagtact caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     4681 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4741 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     4801 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4861 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4921 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4981 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5041 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5101 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5161 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5221 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5281 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5341 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     5401 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5461 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5521 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5581 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5641 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5701 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     5761 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5821 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5881 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     5941 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6001 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6061 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6121 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6181 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6241 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     6301 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6361 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6421 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6481 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6541 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     6601 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6661 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     6721 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     6781 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     6841 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     6901 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     6961 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     7021 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7081 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7141 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7201 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7261 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7321 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7381 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7441 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7501 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7561 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7621 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7681 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7741 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7801 atg
//
