LOCUS       Exported                4783 bp ds-DNA     circular SYN 03-SEP-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4783)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jan 12, 2023 from SnapGene 4.3.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4783
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1..675
                     /organism="pCR8/GW/TOPO"
                     /mol_type="other DNA"
                     /label=source:pCR8/GW/TOPO
     source          683..859
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
     source          join(932..939,2395..2400)
                     /label=Human U6 Promoter
                     /label=Human U6 Promoter(1)
                     /note="/dnas_title=PX335a-U6-chimeric_BB(+85)-CBh-NLS-hSpCs
                     n1( D10A)-NLS"
     source          join(940..966,968..979,1286..1293,1295..1511,1513..1701,
                     1703..1983,1985..2315,2317..2394)
                     /organism="pLenti6/Ubc/V5-DEST Gateway (ViraPower)"
                     /mol_type="other DNA"
                     /label=source:pLenti6/Ubc/V5-DEST Gateway (ViraPower)
     source          join(980..1066,1068..1285)
                     /organism="pLenti6/Ubc/V5-DEST Gateway (ViraPower)"
                     /mol_type="other DNA"
                     /label=source:pLenti6/Ubc/V5-DEST Gateway (ViraPower)
                     /label=source:pLenti6/Ubc/V5-DEST Gateway (ViraPower)(1)
     source          2401..2402
                     /label=Human U6 Promoter
                     /note="/dnas_title=PX335a-U6-chimeric_BB(+85)-CBh-NLS-hSpCs
                     n1( D10A)-NLS"
     source          2456..2622
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          2632..2638
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /label=source:synthetic DNA construct(2)
     source          2661..2889
                     /organism="pCR8/GW/TOPO"
                     /mol_type="other DNA"
                     /label=source:pCR8/GW/TOPO
                     /label=source:pCR8/GW/TOPO(1)
     source          2890..4783
                     /organism="pCR8/GW/TOPO"
                     /mol_type="other DNA"
                     /label=source:pCR8/GW/TOPO
                     /label=source:pCR8/GW/TOPO(1)
     terminator      complement(268..295)
                     /label=rrnB_T2_terminator
     terminator      complement(427..470)
                     /label=rrnB_T1_terminator
     misc_feature    522..544
                     /label=M13_pUC_fwd_primer
     misc_feature    537..553
                     /label=M13_forward20_primer
     misc_feature    complement(569..668)
                     /label=attL1
     misc_feature    671..675
                     /label=EcoRIHS
     promoter        683..859
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     683..703
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     854..859
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    934..939
                     /label=BbsI
                     /label=BbsI(1)
     misc_feature    complement(join(980..1066,1068..1285))
                     /label=ccdB
     misc_feature    complement(1083..1102)
                     /label=AC_S105:hUbcAttRBGHPolyARSeq
     misc_feature    complement(1232..1251)
                     /label=AC_S98:ccdbFSeq
     misc_feature    join(1492..1511,1513)
                     /label=AC_S104:hUbcAttRBGHPolyARSeq
     CDS             complement(join(1627..1701,1703..1983,1985..2286))
                     /codon_start=1
                     /gene="CAT/CamR"
                     /label=CAT/CamR
                     /label=CAT/CamR(1)
                     /note="ORF frame 2"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFHR
                     SGVNTTTISGSFYTYIRKMWRVTVKTWPISLKGLLRICFSSQPIPG*VSPVLI*TWPIW
                     TTSSPPFSPWANIIRKATRC*CRWRFRFIMPL*WLPCRQNA**ITTVLR*VAGRGV"
     gene            complement(join(1627..1701,1703..1983,1985..2286))
                     /gene="CAT/CamR"
                     /label=CAT/CamR
     misc_feature    complement(1702)
                     /label=pLentiSrcReSeqC->TSymInCamR
     misc_feature    complement(1891..1910)
                     /label=AC_S97:CamRFSeq1
     misc_feature    complement(2316)
                     /label=pLentiSrcReseq.G->T
     promoter        complement(2347..2370)
                     /label=lac_promoter
     misc_feature    2395..2400
                     /label=BbsI
     misc_feature    2403..2438
                     /label=PspCas13b_DR
     misc_feature    2662..2761
                     /label=attL2
     promoter        complement(2779..2797)
                     /label=T7_promoter
     misc_feature    complement(2798..2816)
                     /label=M13_reverse_primer
     gene            3030..4040
                     /gene="spect"
                     /label=spect
                     /label=spect(1)
     CDS             3030..4040
                     /codon_start=1
                     /gene="spect"
                     /label=spect
                     /note="ORF frame 2"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4117..4736
                     /label=pBR322_origin
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgatca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtattcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gaattcaccg gtgagggcct atttcccatg attccttcat atttgcatat
      721 acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt
      781 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
      841 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
      901 tttatatatc ttgtggaaag gacgaaacac cgggtcttcg tcgacctgca gactggctgt
      961 gtataaggga gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
     1021 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggacacc ggcacactgg
     1081 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
     1141 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
     1201 cgggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
     1261 tataggtgta aaccttaaac tgcatttcac cagcccctgt tctcgtcagc aaaagagccg
     1321 ttcatttcaa taaaccgggc gacctcagcc atcccttcct gattttccgc tttccagcgt
     1381 tcggcacgca gacgacgggc ttcattctgc atggttgtgc ttaccagacc ggagatattg
     1441 acatcatata tgccttgagc aactgatagc tgtcgctgtc aactgtcact gtaatacgct
     1501 gcttcatagc atacctcttt ttgacatact tcgggtatac atatcagtat atattcttat
     1561 accgcaaaaa tcagcgcgca aatacgcata ctgttatctg gcttttagta agccggatcc
     1621 agatctttac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     1681 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     1741 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     1801 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     1861 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     1921 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     1981 aaaggtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     2041 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     2101 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     2161 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     2221 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     2281 ctccatttta gcttccttag ctcctgaaaa tctcgacgga tcctaactca aaatccacac
     2341 attatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgcgg ccgcgaagac
     2401 ctgttgtgga aggtccagtt ttgaggggct attacaacca attgggtctc cagatttgat
     2461 gtaagggccc attgatgtaa gggcccattg atgtaagcgc gcattgatgt aagcgcgcat
     2521 tgatgtaagc ccggattgat gtaaccgggc attgatgtaa ggtacgccat tgatgtaagg
     2581 cgtaccattg atgtatcgta cccattgatg taagggtacg tagatctttt tttgttttgg
     2641 cgcgccttaa ttaagaattc gacccagctt tcttgtacaa agttggcatt ataaaaaata
     2701 attgctcatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     2761 gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     2821 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     2881 catgcctcct ctagaccagc caggacagaa atgcctcgac ttcgctgctg cccaaggttg
     2941 ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt
     3001 cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
     3061 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
     3121 ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     3181 gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     3241 taaacatcat gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg
     3301 gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag
     3361 tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc
     3421 ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg
     3481 gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc
     3541 cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc
     3601 ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag
     3661 caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc
     3721 ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg
     3781 actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag
     3841 taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg
     3901 cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc
     3961 gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca
     4021 ccaaggtagt cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt
     4081 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4141 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4201 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4261 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4321 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4381 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4441 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4501 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     4561 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4621 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4681 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4741 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
