LOCUS       Exported                4791 bp ds-DNA     circular SYN 01-JUN-2016
DEFINITION  gRNA_Cloning Vector.
ACCESSION   .
VERSION     .
KEYWORDS    gRNA_tdtomato reporter1 SP-2x
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4791)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 1, 2016 from SnapGene 3.1.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4791
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        143..172
                     /label=lac_promoter
                     /note="lac_promoter"
     misc_feature    186..208
                     /label=M13_pUC_rev_primer
                     /note="M13_pUC_rev_primer"
     promoter        207..225
                     /label=M13_reverse_primer
                     /note="M13_reverse_primer promoter"
     CDS             217..336
                     /codon_start=1
                     /label=ORF frame 1
                     /note="ORF frame 1"
                     /translation="MTMITPSYLGDTIEYSSYASSLVPSSDPLVTAASVLEFAL"
     misc_feature    239..256
                     /label=Sp6_primer
                     /note="Sp6_primer"
     misc_feature    complement(337..360)
                     /label=Reverse Primer
                     /note="Reverse Primer"
     misc_feature    complement(414..614)
                     /label=SV40 polyA
                     /note="SV40 polyA"
     misc_feature    complement(698..1399)
                     /label=dTomato
                     /note="dTomato"
     misc_feature    complement(1400..1423)
                     /label=8AA Linker: SGGGLGST
                     /note="8AA Linker: SGGGLGST"
     misc_feature    complement(1424..1426)
                     /label=ScaI
                     /note="ScaI"
     misc_feature    complement(1427..1450)
                     /label=FLAG Tag
                     /note="FLAG Tag"
     misc_feature    complement(1451..1453)
                     /label=Start
                     /note="Start"
     misc_feature    complement(1471..1530)
                     /label=minCMV
                     /note="minCMV"
     misc_feature    complement(1536..1558)
                     /label=AAVS1-gT1
                     /note="AAVS1-gT1"
     misc_feature    complement(1559..1561)
                     /label=ScaI(1)
                     /note="ScaI(1)"
     misc_feature    complement(1562..1579)
                     /label=ISceI DBD
                     /note="ISceI DBD"
     CDS             1586..1664
                     /codon_start=1
                     /label=ORF frame 1(3)
                     /note="ORF frame 1(3)"
                     /translation="MGEFCRYPSHWRPLEHASRGPNSPYS"
     promoter        complement(1656..1664)
                     /label=T7_promoter
                     /note="T7_promoter"
     misc_feature    complement(1665..1687)
                     /label=AAVS1-gT1(1)
                     /note="AAVS1-gT1(1)"
     CDS             1688..2160
                     /codon_start=1
                     /label=ORF frame 1(4)
                     /note="ORF frame 1(4)"
                     /translation="MRITIHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPI
                     ALPNSCAAYTYGSLRFTPIKERAVIVCLWMYRVILLTRRGDGW*SPWPVHVCCQIKSPV
                     NFTRWCISGMKAGA**PPIWPVCRSPLSGKKWLISATAKMTSKTPLT*CSGEY"
     promoter        complement(1688..1697)
                     /label=T7_promoter(1)
                     /note="T7_promoter(1) promoter"
     misc_feature    1701..1844
                     /label=lacZ_a
                     /note="lacZ_a"
     promoter        complement(1704..1720)
                     /label=M13_forward20_primer
                     /note="M13_forward20_primer promoter"
     misc_feature    complement(1713..1735)
                     /label=M13_pUC_fwd_primer
                     /note="M13_pUC_fwd_primer"
     misc_feature    1867..2160
                     /label=ccdB
                     /note="ccdB"
     promoter        2371..2420
                     /label=NEOKAN_promoter
                     /note="NEOKAN_promoter"
     CDS             2509..3303
                     /codon_start=1
                     /label=ORF frame 1(1)
                     /note="ORF frame 1(1)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            2512..3300
                     /gene="NeoR/KanR"
                     /label=NeoR/KanR
                     /note="NeoR/KanR"
     CDS             complement(2818..3351)
                     /codon_start=1
                     /label=ORF frame 1(2)
                     /note="ORF frame 1(2)"
                     /translation="MRKEKIPHQEIVSVNNSEELVKKAIEGDALRIGSGDTVKHEEAVS
                     PFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILAVGH
                     ARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQPPHC
                     ISHDGYFLGRSKVR"
     promoter        3437..3465
                     /label=AmpR_promoter
                     /note="AmpR_promoter"
     misc_feature    3510..3884
                     /label=sh_ble
                     /note="sh_ble"
     gene            3510..3881
                     /gene="bleo"
                     /label=bleo
                     /note="bleo"
     rep_origin      4006..4625
                     /label=pBR322_origin
                     /note="pBR322_origin"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gccctttatt accgcctttg agtgagctga
      361 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagctcgagg
      421 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
      481 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
      541 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
      601 tatgatcctc tagagatatc gtcgacaagc ttatcgatgc ggccgcgcta gcacgcgtca
      661 gctgactaga ggatccccgg gtaccgagct cgaattctta cttgtacagc tcgtccatgc
      721 cgtacaggaa caggtggtgg cggccctcgg agcgctcgta ctgttccacg atggtgtagt
      781 cctcgttgtg ggaggtgatg tccagcttgg tgtccacgta gtagtagccg ggcagttgca
      841 cgggcttctt ggccatgtag atggtcttga actccaccag gtagtggccg ccgtccttca
      901 gcttcagggc ctggtggatc tcgcccttca gcacgccgtc gcgggggtac aggcgctcgg
      961 tggaggcctc ccagcccatg gtcttcttct gcattacggg gccgtcgggg gggaagttgg
     1021 tgccgcgcat cttcaccttg tagatcagcg tgccgtcctg cagggaggag tcctgggtca
     1081 cggtcaccag accgccgtcc tcgaagttca tcacgcgctc ccacttgaag ccctcgggga
     1141 aggacagctt cttgtaatcg gggatgtcgg cggggtgctt cacgtacgcc ttggagccgt
     1201 acatgaactg gggggacagg atgtcccagg cgaagggcag ggggccgccc ttggtcacct
     1261 tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga
     1321 actcgtggcc gttcatggag ccctccatgc gcaccttgaa gcgcatgaac tctttgatga
     1381 cctcctcgcc cttgctcacg gtggaaccca gtccgccacc ggaagtttta tcgtcgtcat
     1441 ccttgtagtc catggtggcg aattctccag gcgatctgac ggttcactaa acgagctctg
     1501 cttatatagg cctcccaccg tacacgccta cctcgcccca ctgtggggtg gaggggacac
     1561 tattaccctg ttatccctac gtcgcaaggg cgaattctgc agatatccat cacactggcg
     1621 gccgctcgag catgcatcta gagggcccaa ttcgccctat agtgccccac tgtggggtgg
     1681 aggggacagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     1741 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     1801 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctata cgtacggcag
     1861 tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt
     1921 gatattattg acacgccggg gcgacggatg gtgatccccc tggccagtgc acgtctgctg
     1981 tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc
     2041 atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat
     2101 ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa
     2161 atgtcaggca tgagattatc aaaaaggatc ttcacctaga tccttttcac gtagaaagcc
     2221 agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg
     2281 gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta
     2341 gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt
     2401 aaggttggga agccctgcaa agtaaactgg atggctttct cgccgccaag gatctgatgg
     2461 cgcaggggat caagctctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     2521 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2581 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2641 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     2701 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     2761 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     2821 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     2881 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     2941 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3001 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     3061 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3121 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3181 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3241 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3301 tgaattatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     3361 atttcacacc gcatacaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3421 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3481 ttcaataata gcacgtgagg agggccacca tggccaagtt gaccagtgcc gttccggtgc
     3541 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     3601 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     3661 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     3721 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     3781 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     3841 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctaaaacttc
     3901 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3961 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4141 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4621 acgcggcctt tttacggttc ctgggctttt gctggccttt tgctcacatg ttctttcctg
     4681 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4741 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
