LOCUS       Exported                8907 bp DNA     circular SYN 30-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8907)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 30, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8907
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          2881..3039
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
     misc_feature    263..826
                     /label=pU6 (pMJ177) 5' CMV promoter IE
     misc_feature    complement(766..775)
                     /label=GC Box
     misc_feature    844..1024
                     /label=HIV-1_5_LTR
     misc_feature    complement(1601..1607)
                     /label=TATA box
     misc_feature    1695..1928
                     /label=RRE
     misc_feature    2459..2473
                     /label=cPPT
     promoter        2631..2871
                     /label=hU6 promoter
     misc_feature    2880..3062
                     /label=CasMINI sgRNA_#2 (sgNT)
     misc_feature    3134..4095
                     /label=EF1Alpha
     misc_feature    4097..4251
                     /label=EF1Alpha
     misc_feature    4320..4976
                     /label=puro-t2A (no BsmBI)
     misc_feature    4536..4542
                     /label=RR47
     misc_feature    4537..4542
                     /label=RR48
     misc_feature    4914..4976
                     /label=T2A
     misc_feature    4989..5684
                     /label=BFP (no BsmBI)
     misc_recomb     5707..5740
                     /label=LoxP recombination site
     misc_feature    5786..6386
                     /label=wPRE
     misc_feature    6576..6590
                     /label=cPPT
     misc_feature    6913..7093
                     /label=HIV-1_5_LTR
     rep_origin      complement(7137..7819)
                     /direction=LEFT
                     /label=ColE1 origin
     misc_feature    complement(7140..7759)
                     /label=pBR322_origin
     misc_feature    complement(7164..7173)
                     /label=GC Box
     CDS             complement(7917..8576)
                     /label=AmpR
     misc_feature    complement(8816..8844)
                     /label=AmpR promoter
ORIGIN
        1 gcttaagcgg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct
     1561 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     1621 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     1681 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     1741 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     1801 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     1861 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     1921 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     1981 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     2041 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     2101 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     2161 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     2221 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     2281 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     2341 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     2401 ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa
     2461 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     2521 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     2581 ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctaga gagggcctat
     2641 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa
     2701 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     2761 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     2821 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     2881 ggcttcactg ataaagtgga gaaccgcttc accaaaagct gtcccttagg ggattagaac
     2941 ttgagtgaag gtgggctgct tgcatcagcc taatgtcgag aagtgctttc ttcggaaagt
     3001 aaccctcgaa acaaattcat ttgaatgaag gaatgcaacc gaatacgccc acgcgatggg
     3061 tattttttta agcttgggcc gctcgaggga tccattaggc ggccgcgtgg ataaccgtat
     3121 taccgccatg catgtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     3181 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
     3241 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     3301 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta
     3361 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct
     3421 tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt
     3481 gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag
     3541 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
     3601 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
     3661 ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt
     3721 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
     3781 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
     3841 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
     3901 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa
     3961 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
     4021 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     4081 ctcgattagt tctcgcgctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     4141 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     4201 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     4261 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgagctag cccaccatga
     4321 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     4381 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     4441 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4501 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4561 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     4621 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     4681 cgtggttcct ggccaccgtc ggcgtttctc cagaccacca gggcaagggt ctgggcagcg
     4741 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     4801 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     4861 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccggatcgg
     4921 gagagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccaccgg
     4981 tcgccaccag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     5041 tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     5101 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     5161 ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact
     5221 tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     5281 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     5341 tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg
     5401 gctgggaggc cttcaccgag actctgtatc ctgctgacgg cggcctggaa ggcagaaacg
     5461 acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacatcaag accacatata
     5521 gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac
     5581 tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg
     5641 ccagatactg cgacctccct agcaaactgg ggcacaagct taattaagaa ttcgtcgagg
     5701 gacctaataa cttcgtatag catacattat acgaagttat acatgtttaa gggttccggt
     5761 tccactaggt acaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     5821 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     5881 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     5941 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     6001 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     6061 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     6121 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     6181 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     6241 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     6301 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     6361 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgatcgaga cctagaaaaa
     6421 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa
     6481 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg
     6541 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     6601 ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc
     6661 tacttccctg attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt
     6721 ggatggtgct acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga
     6781 gagaacaccc gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa
     6841 gtattagagt ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat
     6901 ccggactgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6961 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     7021 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7081 aaaatctcta gcagcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7141 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7201 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7261 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7321 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7381 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7441 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7501 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7561 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7621 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7681 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7741 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7801 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7861 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7921 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7981 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8041 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8101 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8161 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8221 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8281 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8341 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8401 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8461 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8521 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8581 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8641 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8701 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8761 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8821 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8881 acatttcccc gaaaagtgcc acctgac
//
