LOCUS       Exported                4836 bp ds-DNA     circular SYN 05-MAY-2021
DEFINITION  pUC ShDon2-2300.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4836)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 5, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4836
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(231..264)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     terminator      279..350
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        396..500
                     /gene="bla"
                     /label=AmpR promoter
     CDS             501..1361
                     /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      1532..2120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2408..2620
                     /label=Tn RE
                     /note="Tn Right-end"
     terminator      2638..2732
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      2835..2921
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2969..3016
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(3124..3918)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    4305..4352
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    4408..4617
                     /label=Tn LE
                     /note="Tn Left-end"
     terminator      4786..4813
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    complement(join(4813..4836,1..264))
                     /label=sgRNA 2300
                     /note="sgRNA targeting (24bp) eyfp (yellow fluoresence 
                     protein gene)"
ORIGIN
        1 ctttcaaccc atttagggtt cccccttgat cgggggtgat tcagtagcca ctacagcagt
       61 acatccgcgc ccttattgct gggagcgcac ctatcctgtc tattgacagc agcatcaggg
      121 tgagcagcta ccagggtcag aaagcaagac tgtatttata cagtcaaact aaccctcatt
      181 taacaaaatt cagaggctgc aagcagcatg aattgcggcg ctattaatat gctagcatta
      241 tacctaggac tgagctagct gtcagggtct ctctagtaga gagcgttcac cgacaaacaa
      301 cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggag
      361 atcttttacc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      421 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      481 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      541 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      601 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      661 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      721 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      781 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      841 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      901 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      961 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1021 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     1081 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1141 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1201 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1261 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1321 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1381 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1441 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1501 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1561 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1621 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1681 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1741 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1801 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1861 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1921 agctttgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1981 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2041 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2101 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2161 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2221 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2281 cagtgagcga ggaagcggac agggaagagc gcaccggctg gtaacaggat tagcagagcg
     2341 aggcagattg tactgagagt gcaccataca aaaccattgt agagaggtag cattgctacg
     2401 tctctactgt acagtgacta attatatgtc gttgtgacaa attattgtca tcagtaaaat
     2461 ccttatacag tatagattat agcgctttgg cagttttagc ataacctctt tgcagtgaca
     2521 aaatagatgt cgttgtccgt gattgtgaca aattagctgt cgctttgcaa gataggaaaa
     2581 agcttttgtg tattttcata atgacaaatt gactgtcgcc ttaattagct gagcttggac
     2641 tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg
     2701 gttgccgccg ggcgtttttt attggtgaga atccaagcta gactgcgatg agtggcaggg
     2761 cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct
     2821 ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     2881 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgcta ggagcttgcg
     2941 gcccggacga tctcgaattg gggatcttga agtacctatt ccgaagttcc tattctctag
     3001 aaagtatagg aacttcagag cgcttttgaa gctgatgtgc ttaaaaactt actcaatgga
     3061 ataattctag ataattcact agtgattgga aatgttggat cctcatactc ttcctttttc
     3121 aattcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     3181 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     3241 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     3301 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     3361 cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg
     3421 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     3481 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     3541 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     3601 gtgggatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     3661 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     3721 ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     3781 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     3841 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     3901 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc
     3961 cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc
     4021 aaccttacca gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc
     4081 ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
     4141 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     4201 cggactggct ttctacgtgt tccgcttcct ttagcagccc ttgcgccatg catttgttaa
     4261 atcgaattat ccattgctgt tgacaaaggg aatcagggga tcttgaagtt cctattccga
     4321 agttcctatt ctctagaaag tataggaact tcagagcgct tttgaagcta attcgagatc
     4381 atgcatgagc tcactagtgg atccccctca gtaactatta aacttagggg tgggttgaaa
     4441 gcaagtcctt ttatccgctt gttttaattg ctttgtataa taattgcaga gcatattata
     4501 ttgatgacat ttaatttgtc atcaattaat taagcaacgc tgatgggtca cgacgacaat
     4561 taaatagtca caatgacatt aatctgtcac cgacgacaga taatttgtca ctgtacacta
     4621 cgccttttgt ggagatgtct aataaactaa aaatttttta ctttccgctg cataaccctg
     4681 cttcggggtc attgacgaaa gggcctcgtg atctcaggcc gcctaggccg gtttcccgac
     4741 tggaaagcgg ggttccgcgc acatttcccc gaaaagtgcc acctgcgcag aaaggcccac
     4801 ccgaaggtga gccctgcagt gcttcgcccg ctaccc
//
