LOCUS       Exported                4657 bp ds-DNA     circular SYN 22-DEC-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4657)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jan 12, 2023 from SnapGene 4.3.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4657
                     /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          2506..2512
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /label=source:synthetic DNA construct(2)
     source          2535..2763
                     /organism="pCR8/GW/TOPO"
                     /mol_type="other DNA"
                     /label=source:pCR8/GW/TOPO
                     /label=source:pCR8/GW/TOPO(1)
     source          2764..4657
                     /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
     gene            complement(join(1627..1701,1703..1983,1985..2286))
                     /gene="CAT/CamR"
                     /label=CAT/CamR
     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"
     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    2536..2635
                     /label=attL2
     promoter        complement(2653..2671)
                     /label=T7_promoter
     misc_feature    complement(2672..2690)
                     /label=M13_reverse_primer
     CDS             2904..3914
                     /codon_start=1
                     /gene="spect"
                     /label=spect
                     /note="ORF frame 2"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            2904..3914
                     /gene="spect"
                     /label=spect
                     /label=spect(1)
     rep_origin      3991..4610
                     /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 atgtaagatc tttttttgtt ttggcgcgcc
     2521 ttaattaaga attcgaccca gctttcttgt acaaagttgg cattataaaa aataattgct
     2581 catcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     2641 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     2701 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc
     2761 tcctctagac cagccaggac agaaatgcct cgacttcgct gctgcccaag gttgccgggt
     2821 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     2881 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     2941 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     3001 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     3061 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     3121 tcatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     3181 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     3241 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     3301 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     3361 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     3421 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     3481 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     3541 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     3601 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     3661 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     3721 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     3781 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     3841 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     3901 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     3961 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4021 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4081 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4141 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4201 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4261 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4321 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4381 gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4441 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4501 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4561 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4621 ggttcctggc cttttgctgg ccttttgctc acatgtt
//
