LOCUS       Exported                3588 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 3588)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 10, 2022 from SnapGene 5.2.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3588
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     protein_bind    39..60
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        75..105
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    113..129
                     /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     137..153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    139..157
                     /label=M13R
                     /note="M13R"
     CDS             149..266
                     /codon_start=1
                     /label=LacZ
                     /translation="MTMITPSSKLTLTKGNKSWYRAPPRGRRYR*A*YRIPAA"
     promoter        174..192
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    204..266
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     221..237
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    274..751
                     /label=promoter of Chr1 U6 AAEL017702
                     /note="promoter of Chr1 U6 AAEL017702"
     misc_feature    687..699
                     /label=Conserved atccatcgctaga
     misc_feature    721..726
                     /label=TATA
     misc_feature    748..769
                     /label=clone target linkers here
                     /note="clone target linkers here"
     misc_feature    770..855
                     /label=pX330 chimeric guide RNA scaffold
                     /note="pX330 chimeric guide RNA scaffold"
     misc_feature    770..780
                     /label=CRISPR  end
                     /note="CRISPR  end"
     misc_feature    775..805
                     /label=mutations to optimize gRNA according to
                     /note="mutations to optimize gRNA according to Dang et al."
     misc_feature    790..855
                     /label=tracer
                     /note="tracer"
     misc_feature    complement(839..855)
                     /label=EM334
                     /note="EM334"
     misc_feature    856..860
                     /label=U6 Aedes - AAEL017702
     CDS             898..1306
                     /codon_start=1
                     /label=LacZ
                     /translation="GIH*F*SGRHRGGAPIRPIVSRITIHWPSFYNVVTGKTLALPNLI
                     ALQHIPLSPAGVIAKRPAPIALPNSCAA*MANGTRPVAAH*ARRVWWLRAA*PLHLPAP
                     *RPLLSLSSLPFSPRSPAFPVKL*IGGSL*G"
     misc_feature    898..942
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     complement(902..918)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(951..969)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(976..992)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    976..992
                     /label=M13F
                     /note="M13F"
     rep_origin      1134..1589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1615..1719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1720..2580
                     /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      2751..3339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
       61 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      121 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct cgaaattaac
      181 cctcactaaa gggaacaaaa gctggtaccg ggccccccct cgaggtcgac ggtatcgata
      241 agcttgatat cgaattcctg cagcccggtc tcattccata gtaaatatac tacaaataat
      301 attaatgttc catgaaaaaa ggagtaagag tctggtaacc ctagtgcacg caaatatctc
      361 gcgggcatat ttggttgctg aggtatattt atatttgaac gccatgagaa aaagcggaag
      421 aaattggctc atggccgatt ttaaggatat ttaaaaattg tacaatgtac atataattaa
      481 acatccgttc cttcaatgtg ttcttttttt taagcgtgtg ttaaaagttt gctctggtgg
      541 tgaattcacg ctctacccgt tcaggcagca ttcatcgaaa agccctatct gctcgcacac
      601 atttacaaaa tgctgattgc gttgtgtgct gaatgggtca ctcgtccgtc actgcttgct
      661 gtgtacactg tacagttacg cagtctgtgc atcgctagaa tcatatttac ggaaaagtat
      721 tatatatacc caatgcgttg ctcatcggtt gcagtcttcg agaagacctg tttcagagct
      781 atgctggaaa cagcatagca agttgaaata aggctagtcc gttatcaact tgaaaaagtg
      841 gcaccgagtc ggtgcttttt ttgtggtttt attattcgat aattgtggat agagaccggg
      901 ggatccacta gttctagagc ggccgccacc gcggtggagc tccaattcgc cctatagtga
      961 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     1021 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     1081 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     1141 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1201 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1261 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1321 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1381 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1441 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1501 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1561 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     1621 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1681 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1741 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1801 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1861 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1921 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1981 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2041 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2101 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2161 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2221 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2281 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2341 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2401 tttattgctg ataaatctgg agccggtgag cgtggatctc gcggtatcat tgcagcactg
     2461 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2521 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2581 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2641 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2701 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2761 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2821 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2881 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2941 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3001 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3061 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3121 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3181 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3241 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3301 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3361 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3421 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3481 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3541 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacagg
//
