LOCUS       pTrans_9xD3B_CLu        7384 bp    DNA     circular UNA 06-JUL-2015
DEFINITION  CRISPR-Display Cypridina Luciferase-2A-Venus YFP "normalizer,"
            transient format.
ACCESSION   urn.local...1435876244590.78
VERSION     urn.local...1435876244590.78
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     gene            complement(1..861)
                     /gene="bla"
                     /label="bla gene"
     CDS             complement(1..861)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="bla CDS"
     sig_peptide     complement(793..861)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
                     /label="bla signal peptide"
     -10_signal      complement(905..910)
                     /note="bla promoter (counter-clockwise); GAGACA;
                     transcript start 2548 (complementary strand)"
                     /label="-10 signal"
     -35_signal      complement(926..931)
                     /note="bla promoter (counter-clockwise); TTCAAA"
                     /label="-35 signal"
     source          order(1062..>1473,<6193..1061)
                     /organism="Cloning vector pNEB193"
                     /mol_type="other DNA"
                     /label="source Cloning vector pNEB193"
     primer_bind     1413..1436
                     /created_by="User"
                     /label="M13F(-47) From Genewiz"
     primer_bind     1419..1436
                     /created_by="User"
                     /label="M13(-41)F From Genewiz"
     primer_bind     1439..1456
                     /created_by="User"
                     /label="M13F(-21) From Genewiz"
     misc_feature    1493..1512
                     /created_by="User"
                     /label="D3B_gRNA_Binding_Site"
     misc_feature    1526..1545
                     /created_by="User"
                     /label="D3B_gRNA_Binding_Site"
     misc_feature    1564..1583
                     /created_by="User"
                     /label="D3B_gRNA_Binding_Site"
     misc_feature    1597..1616
                     /created_by="User"
                     /label="D3B_gRNA_Binding_Site"
     misc_feature    1640..1659
                     /created_by="User"
                     /label="D3B_gRNA_Binding_Site"
     misc_feature    1673..1692
                     /created_by="User"
                     /label="D3B_gRNA_Binding_Site"
     misc_feature    1711..1730
                     /created_by="User"
                     /label="D3B_gRNA_Binding_Site"
     misc_feature    1744..1763
                     /created_by="User"
                     /label="D3B_gRNA_Binding_Site"
     misc_feature    1777..1796
                     /created_by="User"
                     /label="D3B_gRNA_Binding_Site"
     promoter        1821..1880
                     /gene="minCMV"
                     /modified_by="User"
                     /label="minCMVPro"
     misc_feature    1898..1907
                     /created_by="User"
                     /modified_by="User"
                     /label="Kozak_Sequence"
     sig_peptide     1904..1957
                     /created_by="User"
                     /label="CLuc Signal Peptide"
     CDS             1904..3562
                     /created_by="User"
                     /modified_by="User"
                     /label="Cypridina_Luciferase"
     CDS             3563..3577
                     /created_by="User"
                     /modified_by="User"
                     /label="Linker"
     CDS             3578..3640
                     /created_by="User"
                     /modified_by="User"
                     /label="2A_Self-Cleaving_Peptide"
     CDS             3641..3646
                     /created_by="User"
                     /modified_by="User"
                     /label="Linker"
     CDS             3647..4363
                     /created_by="User"
                     /modified_by="User"
                     /label="Venus YFP"
     misc_feature    complement(3693..3713)
                     /created_by="User"
                     /label="EGFP-N Prtimer from Genewiz"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     4300..4321
                     /created_by="User"
                     /label="EGFP Primer from Genewiz"
     CDS             4364..4372
                     /created_by="User"
                     /label="Linker"
     CDS             4373..4429
                     /created_by="User"
                     /modified_by="User"
                     /label="P2A_Self_Cleaving_Peptide"
     CDS             4430..4435
                     /created_by="User"
                     /label="Linker"
     CDS             4436..5035
                     /created_by="User"
                     /label="Puromycin_Resistance_Gene"
     misc_feature    5057..5654
                     /created_by="User"
                     /modified_by="User"
                     /label="WPRE"
     terminator      5755..5989
                     /created_by="User"
                     /modified_by="User"
                     /label="deltaU3/3' LTR"
     misc_feature    5755..5808
                     /created_by="User"
                     /modified_by="User"
                     /label="deltaU3"
     LTR             5809..5989
                     /created_by="User"
                     /label="3' LTR"
     polyA_signal    6061..6192
                     /created_by="User"
                     /label="SV40 PA signal"
     misc_feature    complement(6098..6117)
                     /created_by="User"
                     /label="SV40 pA Primer from Genewiz"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(6231..6247)
                     /created_by="User"
                     /label="M13Reverse_Primer"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(6280..6285)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(6304..6309)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(6329..6341)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(6626..7214)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(6626..7178)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      7032..7037
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      7054..7059
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        7068..7175
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(7188..7193)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(7209..7214)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
                     /label="-35 signal"
ORIGIN      
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     1021 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     1081 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     1141 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     1201 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     1261 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     1321 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     1381 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     1441 taaaacgacg gccagtgaat tcgagctcgg taccttctcg agctttgcgg taatctagat
     1501 cgcccgtccc ctaggcaaag ctctaatcta gatcgcccgt cccctaggaa aggaagcagc
     1561 cacatctaga tcgcccgtcc cctaggataa cgttagatct agatcgcccg tcccctaggg
     1621 tgaagtagtc tttgcggtaa tctagatcgc ccgtccccta ggcaaagctc taatctagat
     1681 cgcccgtccc ctaggaaagg aagcagccac atctagatcg cccgtcccct aggataacgt
     1741 tagatctaga tcgcccgtcc cctagggtga agtagtatct agatcgcccg tcccctaggc
     1801 cggatccacg tatgtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt
     1861 ttagtgaacc gtcagatcgc ctggaggtac cgccaccgcc accatgaaga ccttaattct
     1921 tgccgttgca ttagtctact gcgccactgt tcattgccag gactgtcctt acgaacctga
     1981 tccaccaaac acagttccaa cttcctgtga agctaaagaa ggagaatgta ttgatagcag
     2041 ctgtggcacc tgcacgagag acatactatc agatggactg tgtgaaaata aaccaggaaa
     2101 aacatgttgc cgaatgtgtc agtatgtaat tgaatgcaga gtagaggccg caggatggtt
     2161 tagaacattc tatggaaaga gattccagtt ccaggaacct ggtacatacg tgttgggtca
     2221 aggaaccaag ggcggcgact ggaaggtgtc catcaccctg gagaacctgg atggaaccaa
     2281 gggggctgtg ctgaccaaga caagactgga agtggctgga gacatcattg acatcgctca
     2341 agctactgag aatcccatca ctgtaaacgg tggagctgac cctatcatcg ccaacccgta
     2401 caccatcggc gaggtcacca tcgctgttgt tgagatgcca ggcttcaaca tcaccgtcat
     2461 tgagttcttc aaactgatcg tgatcgacat cctcggagga agatctgtaa gaatcgcccc
     2521 agacacagca aacaaaggaa tgatctctgg cctctgtgga gatcttaaaa tgatggaaga
     2581 tacagacttc acttcagatc cagaacaact cgctattcag cctaagatca accaggagtt
     2641 tgacggttgt ccactctatg gaaatcctga tgacgttgca tactgcaaag gtcttctgga
     2701 gccgtacaag gacagctgcc gcaaccccat caacttctac tactacacca tctcctgcgc
     2761 cttcgcccgc tgtatgggtg gagacgagcg agcctcacac gtgctgcttg actacaggga
     2821 gacgtgcgct gctcccgaaa ctagaggaac ctgcgttttg tctggacata ctttctacga
     2881 tacatttgac aaagcaagat accaattcca gggtccctgc aaggagattc ttatggccgc
     2941 cgactgtttc tggaacactt gggatgtgaa ggtttcacac aggaatgttg actcttacac
     3001 tgaagtagag aaagtacgaa tcaggaaaca atcgactgta gtagaactca ttgttgatgg
     3061 aaaacagatt ctggttggag gagaagccgt gtccgtcccg tacagctctc agaacacttc
     3121 catctactgg caagatggtg acatactgac tacagccatc ctacctgaag ctctggtggt
     3181 caagttcaac ttcaagcaac tgctcgtcgt acatattaga gatccattcg atggtaagac
     3241 ttgcggtatt tgcggtaact acaaccagga tttcagtgat gattcttttg atgctgaagg
     3301 agcctgtgat ctgaccccca acccaccggg atgcaccgaa gaacagaaac ctgaagctga
     3361 acgactctgc aatagtctct tcgccggtca aagtgatctt gatcagaaat gtaacgtgtg
     3421 ccacaagcct gaccgtgtcg aacgatgcat gtacgagtat tgcctgaggg gacaacaggg
     3481 tttctgtgac cacgcatggg agttcaagaa agaatgctac ataaagcatg gagacaccct
     3541 agaagtacca gatgaatgca aaggcagcac cagcggcggc agtggagagg gcagaggaag
     3601 tctgctaaca tgcggtgacg tcgaggagaa tcctggccca ggcagtatgg tgagcaaggg
     3661 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     3721 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     3781 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     3841 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     3901 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3961 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     4021 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     4081 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     4141 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
     4201 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     4261 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     4321 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcagcg gtgctacgaa
     4381 cttcagtctc ctgaaacagg ctggggatgt ggaggagaac cccggcccag gcagcatgac
     4441 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     4501 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     4561 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     4621 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     4681 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     4741 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     4801 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     4861 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     4921 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     4981 cgagtgcccg aaggaccgcg cgacctggtg catgacccgc aagcccggtg cctagtaacg
     5041 taccggttag taatgatcga caatcaacct ctggattaca aaatttgtga aagattgact
     5101 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     5161 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     5221 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     5281 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     5341 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5401 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     5461 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5521 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5581 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5641 gcctccccgc ctggcgatgg taccgctaag cacaattcga gctcggtacc tttaagacca
     5701 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     5761 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     5821 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     5881 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     5941 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     6001 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     6061 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6121 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6181 tatcatgtct gggtttaaac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg
     6241 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6301 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6361 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6421 gcggggagag gcggtttgcg tattgggccg cttcctcgct cactgactcg ctgcgctcgg
     6481 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6541 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6601 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6661 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6721 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6781 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6841 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6901 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6961 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7021 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     7081 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7141 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7201 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7261 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7321 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7381 acag
//
