LOCUS       Exported                3477 bp ds-DNA     circular SYN 01-JUN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 3477)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 10, 2022 from SnapGene 5.2.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3477
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(286..694)
                     /codon_start=1
                     /label=LacZ
                     /translation="GIH*F*SGRHRGGAPIRPIVSRITIHWPSFYNVVTGKTLALPNLI
                     ALQHIPLSPAGVIAKRPAPIALPNSCAA*MANGTRPVAAH*ARRVWWLRAA*PLHLPAP
                     *RPLLSLSSLPFSPRSPAFPVKL*IGGSL*G"
     misc_feature    600..616
                     /label=M13F
                     /note="M13F"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    650..694
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     674..690
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    702..1022
                     /label=pU6 promoter Chr3 AAEL017905 Konet2007
                     /note="pU6 promoter Chr3 AAEL017905 Konet2007"
     misc_feature    982..995
                     /label=conserved
     misc_feature    1016..1021
                     /label=TATA
     misc_feature    1023..1047
                     /label=this section from AAEL017774 U6
                     /note="this section from AAEL017774 U6"
     misc_feature    1045..1066
                     /label=clone target linkers here
                     /note="clone target linkers here"
     misc_feature    1048
                     /label=promoter of Chr1 U6 AAEL017702
                     /note="promoter of Chr1 U6 AAEL017702"
     misc_feature    1067..1152
                     /label=pX330 chimeric guide RNA scaffold
                     /note="pX330 chimeric guide RNA scaffold"
     misc_feature    1067..1077
                     /label=CRISPR  end
                     /note="CRISPR  end"
     misc_feature    1072..1102
                     /label=mutations to optimize gRNA according to
                     /note="mutations to optimize gRNA according to Dang et al."
     misc_feature    1087..1152
                     /label=tracer
                     /note="tracer"
     misc_feature    complement(1136..1152)
                     /label=EM334
                     /note="EM334"
     misc_RNA        1153..1157
                     /label=AAEL017905-RA
                     /note="snRNA"
     CDS             complement(1215..1332)
                     /codon_start=1
                     /label=LacZ
                     /translation="MTMITPSSKLTLTKGNKSWYRAPPRGRRYR*A*YRIPAA"
     misc_feature    1215..1277
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     complement(1244..1260)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1289..1307)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    complement(1324..1342)
                     /label=M13R
                     /note="M13R"
     primer_bind     complement(1328..1344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1352..1368
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1376..1406)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1421..1442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1730..2318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2489..3349)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3350..3454)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
      661 cggtggcggc cgctctagaa ctagtggatc ccccggtctc actctaattg gagctgccag
      721 acaaatttga tttgtccgtg cggtatgcat atgtacctac ttacgtctgt cgttttgtct
      781 ccgtttaccg ggagggaaag tctggaaaca tggaaactct atagttgcca ggtagaccat
      841 ctgcctccgt cggctggctg gattccaatt tgaatattgg ctaattggaa gagatggaag
      901 tttttgaatg gatgattgaa taattgaagc gactccgggt acctgtttgt aagctctgca
      961 acagtgccat agattcgtgt cagtccatca ctagaatcaa atcaacttgt acttgatata
     1021 taagagcaga ggcaagagta gtgaaatgca gtcttcgaga agacctgttt cagagctatg
     1081 ctggaaacag catagcaagt tgaaataagg ctagtccgtt atcaacttga aaaagtggca
     1141 ccgagtcggt gcttttttta caaatctatt atgcatgagg tactatgttg gcagagtaat
     1201 gaattccaga gaccgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg
     1261 agggggggcc cggtaccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
     1321 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1381 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1441 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1501 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1561 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1621 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1681 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1741 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1801 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1861 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1921 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1981 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2041 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2101 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2161 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2221 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2281 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2341 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2401 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2461 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2521 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2581 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct
     2641 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2701 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2761 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2821 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2881 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2941 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3001 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3061 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3121 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3181 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3241 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3301 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3361 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3421 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
