LOCUS       pXJ218_AAV_SRT_gRNA_dir 6873 bp ds-DNA     circular     13-MAY-2023
DEFINITION  .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     9Backbone without insert 4.4kb.  Imported using the Genbank
            importer. File name: addgene-plasmid-154889-sequence-306065.gbk
            piggyBac self-reporting transposon.
FEATURES             Location/Qualifiers
     CDS             complement(171..1031)
                     /label="AmpR"
     primer_bind     794..813
                     /label="Amp-R"
     promoter        complement(1032..1095)
                     /label="AmpR promoter"
     misc_feature    1475..1491
                     /label="M13f40-genewiz"
     primer_bind     1480..1502
                     /label="M13/pUC Forward"
     primer_bind     1494..1511
                     /label="M13 Forward"
     primer_bind     1495..1511
                     /label="M13 fwd"
     promoter        1522..1540
                     /label="T7 promoter"
     primer_bind     1522..1541
                     /label="T7"
     repeat_region   1577..1706
                     /label="AAV2 ITR"
     repeat_region   1853..1915
                     /label="piggyBac right (3') inverted repeat"
     misc_feature    2150..2390
                     /label="U6 prom"
     misc_feature    2399..2416
                     /label="empty guide"
     misc_feature    2417..2492
                     /label="gRNA scaffold"
     misc_feature    2449..2469
                     /label="5' kit"
     misc_feature    2493..2514
                     /label="direct capture 1 (10x genomics)"
     promoter        2539..3710
                     /label="EF1Alpha"
     misc_feature    3675..3719
                     /label="fwd overhang"
     mRNA            3720..4418
                     /label="mTagBFP"
                     /note="vntifkey: 54"
     CDS             3720..4418
                     /label="Translation 3720-4418"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCDLPSKLGHKLN"
     misc_feature    4419..4685
                     /label="KASH-tn5confimred"
     CDS             4419..4685
                     /label="Translation 4419-4685"
                     /translation="SGLRSREEEEETDSRMPHLDSPGSSQPRRSFLSRVIRAALPLQLLLLLLLLLACLLPASEDDYSCTQANNFARSFYPMLRYTNGPPPT*"
     misc_feature    4692..4720
                     /label="rev overhang"
     misc_feature    4735..5211
                     /label="hGH polyA signal"
     misc_feature    5224..5243
                     /label="salazar rev1"
     misc_feature    5244..5272
                     /label="AS2_gibson_flanking"
     repeat_region   5532..5566
                     /label="piggyBac left (5') inverted repeat"
     repeat_region   5874..6003
                     /label="AAV2 ITR"
     primer_bind     complement(6131..6148)
                     /label="L4440"
     terminator      6185..6228
                     /label="rrnB T1 terminator"
     rep_origin      complement(6285..6873)
                     /label="ori"
     primer_bind     complement(6365..6384)
                     /label="pBR322ori-F"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga agctaataac aggcctgctg gtaatcgcag gcctttttat
     1141 ttggtgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     1201 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     1261 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     1321 gagatagggt tgagtggccg ctacagggcg ctcccattcg ccattcaggc tgcgcaactg
     1381 ttgggaaggg cgtttcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     1441 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     1501 cgacggccag tgagcgcgac gtaatacgac tcactatagg gcgaattgta ggcggaaggc
     1561 cgtcaaggcc gcatggctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
     1621 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     1681 ggccaactcc atcactaggg gttcctattg acgtcagatc gctaaataat gacgtatgtt
     1741 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     1801 actgcccact tggcagtaca tcaagtgtat catacgtcaa ggccgcattt aaccctagaa
     1861 agataatcat attgtgacgt acgttaaaga taatcatgtg taaaattgac gcatgtgttt
     1921 tatcggtctg tatatcgagg tttatttatt aatttgaata gatattaagt tttattatat
     1981 ttacacttac atactaataa taaattcaac aaacaattta tttatgttta tttatttatt
     2041 aaaaaaaaca aaaactcaaa atttcttcta taaagtaaca aaacttttat cgaattcctg
     2101 cagcccagct tgatcttagg tccctcgacg aattctctag agcacgcgtg agggcctatt
     2161 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
     2221 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
     2281 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
     2341 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccGG
     2401 AAGAGCGAGC TCTTCTGTTT TAGAGCTAGA AATAGCAAGT TAAAATAAGG CTAGTCCGTT
     2461 ATCAACTTGA AAAAGTGGCA CCGAGTCGGT GCGCTTTAAG GCCGGTCCTA GCAAtttttt
     2521 tctagactgc agagctgcGT GCCCGTCAGT GGGCAGAGCG CACATCGCCC ACAGTCCCCG
     2581 AGAAGTTGGG GGGAGGGGTC GGCAATTGAA CCGGTGCCTA GAGAAGGTGG CGCGGGGTAA
     2641 ACTGGGAAAG TGATGTCGTG TACTGGCTCC GCCTTTTTCC CGAGGGTGGG GGAGAACCGT
     2701 ATATAAGTGC AGTAGTCGCC GTGAACGTTC TTTTTCGCAA CGGGTTTGCC GCCAGAACAC
     2761 AGGTAAGTGC CGTGTGTGGT TCCCGCGGGC CTGGCCTCTT TACGGGTTAT GGCCCTTGCG
     2821 TGCCTTGAAT TACTTCCACC TGGCTGCAGT ACGTGATTCT TGATCCCGAG CTTCGGGTTG
     2881 GAAGTGGGTG GGAGAGTTCG AGGCCTTGCG CTTAAGGAGC CCCTTCGCCT CGTGCTTGAG
     2941 TTGAGGCCTG GCCTGGGCGC TGGGGCCGCC GCGTGCGAAT CTGGTGGCAC CTTCGCGCCT
     3001 GTCTCGCTGC TTTCGATAAG TCTCTAGCCA TTTAAAATTT TTGATGACCT GCTGCGACGC
     3061 TTTTTTTCTG GCAAGATAGT CTTGTAAATG CGGGCCAAGA TCTGCACACT GGTATTTCGG
     3121 TTTTTGGGGC CGCGGGCGGC GACGGGGCCC GTGCGTCCCA GCGCACATGT TCGGCGAGGC
     3181 GGGGCCTGCG AGCGCGGCCA CCGAGAATCG GACGGGGGTA GTCTCAAGCT GGCCGGCCTG
     3241 CTCTGGTGCC TGGCCTCGCG CCGCCGTGTA TCGCCCCGCC CTGGGCGGCA AGGCTGGCCC
     3301 GGTCGGCACC AGTTGCGTGA GCGGAAAGAT GGCCGCTTCC CGGCCCTGCT GCAGGGAGCT
     3361 CAAAATGGAG GACGCGGCGC TCGGGAGAGC GGGCGGGTGA GTCACCCACA CAAAGGAAAA
     3421 GGGCCTTTCC GTCCTCAGCC GTCGCTTCAT GTGACTCCAC GGAGTACCGG GCGCCGTCCA
     3481 GGCACCTCGA TTAGTTCTCG cGCTTTTGGA GTACGTCGTC TTTAGGTTGG GGGGAGGGGT
     3541 TTTATGCGAT GGAGTTTCCC CACACTGAGT GGGTGGAGAC TGAAGTTAGG CCAGCTTGGC
     3601 ACTTGATGTA ATTCTCCTTG GAATTTGCCC TTTTTGAGTT TGGATCTTGG TTCATTCTCA
     3661 AGCCTCAGAC AGTGGTTCAA AGTTTTTTTC TTCCATTTCA GGTGTCGTGA gCTAGCACCa
     3721 tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca
     3781 accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     3841 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta
     3901 gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca
     3961 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     4021 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     4081 tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     4141 aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg
     4201 ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca tatagatcca
     4261 agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa
     4321 gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat
     4381 actgcgacct ccctagcaaa ctggggcaca agcttaatTC CGGACTCAGA TCTCGAGAGG
     4441 AGGAGGAGGA GACAGACAGC AGGATGCCCC ACCTCGACAG CCCCGGCAGC TCCCAGCCGA
     4501 GACGCTCCTT CCTCTCAAGG GTGATCAGGG CAGCGCTACC GTTGCAGCTG CTTCTGCTGC
     4561 TGCTGCTGCT CCTGGCCTGC CTGCTACCTG CCTCTGAAGA TGACTACAGC TGCACCCAGG
     4621 CCAACAACTT TGCCCGATCC TTCTACCCCA TGCTGCGGTA CACCAACGGG CCACCTCCCA
     4681 CCTAGgaatt cgatatcaag cttatcgata ccgagcgctg ctcgagagat ctacgggtgg
     4741 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     4801 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     4861 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     4921 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     4981 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     5041 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     5101 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     5161 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     5221 taggtaacca cgtgcggacc gagcgccggc gcgataagct atctataaca agaaaatata
     5281 tatataataa gttatcacgt aagtagaaca tgaaataaca atataattat cgtatgagtt
     5341 aaatcttaaa agtcacgtaa aagataatca tgcgtcattt tgactcacgc ggtcgttata
     5401 gttcaaaatc agtgacactt accgcattga caagcacgcc tcacgggagc tccaagcggc
     5461 gactgagatg tcctaaatgc acagcgacgg attcgcgcta tttagaaaga gagagcaata
     5521 tttcaagaat gcatgcgtca attttacgca gactatcttt ctagggttaa gagctcctga
     5581 gatgcaggta catcccactg atgagtccca aataggacga aacgcgcttc ggtgcgtctg
     5641 ggattccact gctatccacg cggccgcacg gcattacgtt agacgtctag gccacaccac
     5701 cctgaaggcg cctgctcgcc tctgatctgt tgaagctaag cagggtcggt cctggttagt
     5761 acttggatgg gactccgcct ggtaatagcc ggtaccgtag gcttgcatgg tcatagctgt
     5821 ttccttgcgc tggggatgcg gtgggctcta tagagagtcg atcgtgtgat cgaaggaacc
     5881 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     5941 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     6001 cagcatgtag agactgggcc tcatgggcct tccgcctatc actgcccgct ttccagtcgg
     6061 gaaacctgtc gtgccagctg cattaacatg gtcatagctg tttccttgcg tattgggcgc
     6121 tctccgcttc ctcgctcact gactcgctgc gctcggtcgt tcgggtaaag cctggggtgc
     6181 ctaaataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattg agcaaaaggc
     6241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6661 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6721 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6781 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6841 cagcagatta cgcgcagaaa aaaaggatct caa
//