LOCUS       Exported                3487 bp ds-DNA     circular SYN 14-NOV-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 3487)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 10, 2022 from SnapGene 5.2.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3487
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     protein_bind    23..44
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        59..89
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    97..113
                     /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)."
     primer_bind     121..137
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    123..141
                     /label=M13R
                     /note="M13R"
     CDS             133..250
                     /codon_start=1
                     /label=LacZ
                     /translation="MTMITPSSKLTLTKGNKSWYRAPPRGRRYR*A*YRIPAA"
     promoter        158..176
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    188..250
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     205..221
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    258..617
                     /label=U6 prom from Chr3 AAEL017763
                     /note="U6 prom from Chr3 AAEL017763"
     misc_feature    554..567
                     /label=Conserved
     misc_feature    587..592
                     /label=TATA
     misc_feature    615..636
                     /label=clone target linkers here
                     /note="clone target linkers here"
     misc_feature    637..722
                     /label=pX330 chimeric guide RNA scaffold
                     /note="pX330 chimeric guide RNA scaffold"
     misc_feature    637..647
                     /label=CRISPR  end
                     /note="CRISPR  end"
     misc_feature    642..672
                     /label=mutations to optimize gRNA according to
                     /note="mutations to optimize gRNA according to Dang et al."
     misc_feature    657..722
                     /label=tracer
                     /note="tracer"
     misc_feature    complement(706..722)
                     /label=EM334
                     /note="EM334"
     misc_RNA        723..727
                     /label=U6 AAEL017763-RA
                     /note="snRNA"
     CDS             781..1189
                     /codon_start=1
                     /label=LacZ
                     /translation="GIH*F*SGRHRGGAPIRPIVSRITIHWPSFYNVVTGKTLALPNLI
                     ALQHIPLSPAGVIAKRPAPIALPNSCAA*MANGTRPVAAH*ARRVWWLRAA*PLHLPAP
                     *RPLLSLSSLPFSPRSPAFPVKL*IGGSL*G"
     misc_feature    781..825
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     complement(785..801)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(834..852)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    859..875
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(859..875)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1017..1472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1498..1602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1603..2463
                     /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"
     rep_origin      2634..3222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
       61 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      121 caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac
      181 aaaagctggt accgggcccc ccctcgaggt cgacggtatc gataagcttg atatcgaatt
      241 cctgcagccc ggtctcatgt tcgttcttca acacctctcc atggtgataa cggatacggt
      301 ttcattgtca gcatccatcc tccgaaaaat acattacgcc ttgaaatatg caatcgcaaa
      361 cacggatctg tttggaacat ttattttact atgaagagat gcgataggta atatttattt
      421 gagcgtttaa gatactcatt gttctctcaa agaatgtcat tgaaagccaa cgaggtcaaa
      481 tcaaatatta taataaaaag gtcaaagagg actaacttaa agctctcttt atggatagga
      541 aaaaatattt tcgcccatcg ctagaacttt taccgtttcc attgagtata taactaagat
      601 gaatgaggct aattgatgca gtcttcgaga agacctgttt cagagctatg ctggaaacag
      661 catagcaagt tgaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
      721 gcttttttaa gatgcttcgg caattaataa taaggaatgg taaagcattc tctagagacc
      781 gggggatcca ctagttctag agcggccgcc accgcggtgg agctccaatt cgccctatag
      841 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
      901 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
      961 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     1021 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1081 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1141 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1201 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1261 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1321 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1381 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1441 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     1501 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1561 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1621 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1681 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1741 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1801 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1861 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1921 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1981 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2041 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2101 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2161 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2221 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2281 tggtttattg ctgataaatc tggagccggt gagcgtggat ctcgcggtat cattgcagca
     2341 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2401 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2461 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2521 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2581 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2641 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2701 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2761 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2821 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2881 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2941 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3001 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3061 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3121 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3181 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3241 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3301 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3361 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3421 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     3481 tggaaag
//
