LOCUS       pT4-U6-sgRNA-CMV-EGFP   7260 bp ds-DNA     circular     31-JAN-2024
DEFINITION  .
ACCESSION   pVec1
VERSION     pVec1.1
COMMENT     Imported using the Genbank importer. File name:
            pT4-U6-sgRNA-CMV-EGFP.gb Linear. pT4-CMV-GFP with HindIII ->
            InFusion with amplicon (template: pT4-U6-sgRNA) >InF-U6-F
            ctgtgcctttaagcttggtaccgagggcctatttccc >InF-sgRNA-Hind-R
            aatcaatgccaagcttaaaaaagcaccgactcggtgc Ta: 70°C - ext.: 2250bp
            Screen by HindIII -> .7 + .85kb pos. fragments
FEATURES             Location/Qualifiers
     CDS             complement(138..206)
                     /label="LacZ alpha"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     277..294
                     /label="M13-fwd"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     303..323
                     /label="T7"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    347..1347
                     /label="sanger (T7)"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    complement(348..591)
                     /label="pT4 LIR"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          582..618
                     /label="InF-U6-F"
                     /note="sequence: ctgtgcctttaagcttggtaccgagggcctatttccc"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          604..624
                     /label="hU6-f_Microsynth"
                     /note="sequence: GAGGGCCTATTTCCCATGATT"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    604..853
                     /label="U6 promoter"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          773..797
                     /label="qPCR-screen-U6-F2"
                     /note="sequence: tggactatcatatgcttaccgtaac"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          835..852
                     /label="qPCR-screen-U6-F"
                     /note="sequence: tggaaaggacgaaacacc"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    854..2733
                     /label="filler"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(856..876)
                     /label="qPCR-screen-filler-R2"
                     /note="sequence: gtgtgctcatttacaaccgtc"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          complement(1030..1049)
                     /label="qPCR-screen-filler-R"
                     /note="sequence: aagaaaacaacgcgagttac"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    complement(2091..3121)
                     /label="sanger (CMV-rev)"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    2734..2815
                     /label="chimeric RNA backbone 1 (BsmBI)"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(2795..2831)
                     /label="InF-sgRNA-Hind-R"
                     /note="sequence: aatcaatgccaagcttaaaaaagcaccgactcggtgc"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    2822..3201
                     /label="CMV enhancer"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    3202..3405
                     /label="CMV promoter"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    3497..3629
                     /label="chimeric intron"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    3634..3643
                     /label="kozak"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    3640..4359
                     /label="EGFP CDS"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    4366..4414
                     /label="synthetic poly(A) signal"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    4452..4717
                     /label="pT4 RIR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     complement(4748..4767)
                     /label="T3"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     promoter        complement(4750..4768)
                     /label="T3 promoter"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     complement(4785..4805)
                     /label="M13-rev"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    complement(4811..4833)
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     promoter        complement(4837..4867)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    complement(5191..5779)
                     /label="Ori"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    complement(5950..6810)
                     /label="AmpR"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
ORIGIN
        1 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
       61 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      121 attaacgctt acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg
      181 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
      241 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
      301 gcgtaatacg actcactata gggcgaattg gagctagcgt ttaaacctac agttgaagtc
      361 ggaagtttac atacacttaa gttggagtca ttaaaactcg tttttcaact actccacaaa
      421 tttcttgtta acaaacaata gttttggcaa gtcagttagg acatctactt tgtgcatgac
      481 acaagtcatt tttccaacaa ttgtttacag acagattatt tcacttataa ttcactgtat
      541 cacaattcca gtgggtcaga agtgtacata cacgcgcttg actgtgcctt taagcttggt
      601 accgagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
      661 gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
      721 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
      781 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
      841 ggacgaaaca ccggagacgg ttgtaaatga gcacacaaaa tacacatgct aaaatattat
      901 attctatgac ctttataaaa tcaaccaaaa tcttcttttt aataacttta gtatcaataa
      961 ttagaatttt tatgttcctt tttgcaaact tttaataaaa atgagcaaaa taaaaaaacg
     1021 ctagttttag taactcgcgt tgttttcttc acctttaata atagctactc caccacttgt
     1081 tcctaagcgg tcagctcctg cttcaatcat tttttgagca tcttcaaatg ttctaactcc
     1141 accagctgct ttaactaaag cattgtcttt aacaactgac ttcattagtt taacatcttc
     1201 aaatgttgca cctgattttg aaaatcctgt tgatgtttta acaaattcta atccagcttc
     1261 aacagctatt tcacaagctt tcatgatttc ttcttttgtt aataaacaat tttccataat
     1321 acatttaaca acatgtgatc cagctgcttt ttttacagct ttcatgtctt ctaaaactaa
     1381 ttcataattt ttgtctttta atgcaccaat atttaatacc atatcaattt ctgttgcacc
     1441 atctttaatt gcttcagaaa cttcgaatgc ttttgtagct gttgtgcatg cacctagagg
     1501 aaaacctaca acatttgtta ttcctacatt tgtgcctttt aataattctt tacaatagct
     1561 tgttcaatat gaattaacac aaactgttgc aaaatcaaat tcaattgctt catcacataa
     1621 ttgtttaatt tcagctttcg tagcatcttg ttttaataat gtgtgatcta tatatttgtt
     1681 tagtttcatt ttttctccta tatattcatt tttaatttta attctttaat aatttcgtct
     1741 actttaactt tagcgttttg aacagattca ccaacaccta taaaataaat ttttagttta
     1801 ggttcagttc cacttgggcg aacagcaaat catgacttat cttctaaata aaattttagt
     1861 aagtcttgtc ctggcatatt atacattcca tcgatgtagt cttcaacatt aacaacttta
     1921 agtccagcaa tttgagttaa gggtgttgct ctcaatgatt tcattaatgg ttcaattttt
     1981 aatttctttt cttctggttt aaaattcaag tttaaagtga aagtgtaata tgcacccatt
     2041 tctttaaata aatcttctaa atagtctact aatgttttat tttgtttttt ataaaatcaa
     2101 gcagcctctg ctattaatat agaagcttgt attccatctt tatctctagc tgagtcatca
     2161 attacatatc cataactttc ttcataagca aaaacaaaat ttaatccgtt atcttcttct
     2221 ttagcaattt ctctacccat tcatttaaat ccagttaaag tttttacaat attaactcca
     2281 tatttttcat gagcgattct atcacccaaa tcacttgtta caaaacttga atatagagcc
     2341 ggattttttg gaatgctatt taagcgtttt agatttgata attttcaatc aattaaaatt
     2401 ggtcctgttt gatttccatc taatcttaca aaatgaccat catgttttat tgccattcca
     2461 aatctgtcag catctgggtc attcataata ataatatctg catcatgttt aataccatat
     2521 tcaagcggta tttttcatgc aggatcaaat tctggatttg gatttacaac atttttaaat
     2581 gtttcatctt caaatgcatg ctcttcaacc tcaataacgt tatatcctga ttcacgtaat
     2641 atttttgggg taaatttagt tcctgttcca ttaactgcgc taaaaataat ttttaaatct
     2701 tttttagctt cttgctcttt tttgtacgtc tctgttttag agctagaaat agcaagttaa
     2761 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttaagct
     2821 tggcattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     2881 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     2941 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     3001 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     3061 caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc
     3121 tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta catctacgta
     3181 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag
     3241 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     3301 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa
     3361 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gaggtcgttt agtgaaccgt
     3421 cagatcacta gtagctttat tgcggtagtt tatcacagtt aaattgctaa cgcagtcagt
     3481 gctcgactga tcacaggtaa gtatcaaggt tacaagacag gtttaaggag gccaatagaa
     3541 actgggcttg tcgagacaga gaagattctt gcgtttctga taggcaccta ttggtcttac
     3601 tgacatccac tttgcctttc tctccacagg gcggccgcca tggtgagcaa gggcgaggag
     3661 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     3721 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     3781 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     3841 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     3901 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     3961 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     4021 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     4081 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     4141 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     4201 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     4261 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     4321 gccgggatca ctctcggcat ggacgagctg tacaagtaag aattcaataa aatatcttta
     4381 ttttcattac atctgtgtgt tggttttttg tgtgtaattc gagcatctta ccgccattta
     4441 ttcccagatc tttaaacaat ttaaaggcaa tgctaccaaa tactaagcgc gtgtatgtac
     4501 acttctgacc cactgggaat gtgatgaaag aaataaaagc tgaaatgaat cattctctct
     4561 actattattc tgatatttca cattcttaaa ataaagtggt gatcctaact gaccttaaga
     4621 cagggaatct ttactcggat taaatgtcag gaattgtgaa aaagtgagtt taaatgtatt
     4681 tggctaaggt gtatgtaaac ttccgacttc aactgtagtt taaacgggcc cggtacccag
     4741 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     4801 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4861 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4921 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4981 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5041 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5101 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5161 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5221 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5281 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5341 atacctgtcc gcctttctcc cttcgggaac cgtggcgctt tctcatagct cacgctgtag
     5401 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5461 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5521 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5581 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5641 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5701 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5761 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5821 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5881 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5941 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6001 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6061 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6121 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6181 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6241 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6301 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6361 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaact tggccgcagt
     6421 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6481 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6541 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6601 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6661 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6721 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6781 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6841 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6901 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     6961 attaagcgcg gcgggagtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     7021 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     7081 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     7141 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     7201 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
//