LOCUS       Exported                3757 bp ds-DNA     circular SYN 18-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3757)
  AUTHORS   Doudna Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 28, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3757
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(40..94)
                     /label=oBA1714_p15_cmR_part4_gib_R
     primer_bind     complement(69..91)
                     /label=oBA1740_pt4_commonGib_R
     primer_bind     complement(73..94)
                     /label=oBA1651_pBFC0924_Gib_R
     misc_feature    88..120
                     /label=gbBA079
     misc_feature    88..91
                     /label=4>5
     misc_feature    92..116
                     /label=ConS scar
     primer_bind     92..116
                     /label=oBA1097_conS_Scar_check_F
     protein_bind    92..95
                     /label=BsmBI
     primer_bind     complement(99..135)
                     /label=HS_690
     primer_bind     106..142
                     /label=HS_689
     misc_feature    117..120
                     /label=Pt2/2a BsaI Junction Scar
     terminator      121..487
                     /label=TrrnB terminator (BBa_K1893035)
                     /label=TrrnB
     misc_feature    121..124
                     /label=DN
     misc_feature    122
                     /label=a>t removes HindIII
     misc_feature    join(208,221)
                     /label=covariant muts to remove BsaI
     misc_feature    join(390,407)
                     /label=covariant muts to remove BsaI
     terminator      421..466
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(460..487)
                     /label=oBFC2037
     misc_feature    484..487
                     /label=BsmBI_OH
     primer_bind     487..515
                     /label=oBFC1697
     primer_bind     487..515
                     /label=PTet_F
     misc_feature    488..491
                     /label=1>2
     misc_feature    492..1231
                     /label=PCR from pba369
     primer_bind     492..507
                     /label=oBA1632_EJC_pTET_p2_F
     primer_bind     492..506
                     /label=oBFC2038
     primer_bind     492..506
                     /label=oBFC0336
     CDS             complement(499..1125)
                     /label=tetR
     primer_bind     1015..1041
                     /label=KEW.A01
     primer_bind     1066..1088
                     /label=oBFC0419
     primer_bind     complement(1085..1122)
                     /label=oBFC0604
     promoter        1126..1198
                     /label=tet promoter region
     primer_bind     1126..1169
                     /label=oBFC1701
     protein_bind    1147..1165
                     /label=Q1
     primer_bind     complement(1162..1196)
                     /label=oBFC0337
     primer_bind     complement(1162..1196)
                     /label=oBFC0338
     primer_bind     complement(1169..1203)
                     /label=HS_672
     primer_bind     complement(1174..1198)
                     /label=oBFC1704
     primer_bind     1176..1194
                     /label=oBFC1899
     protein_bind    1177..1193
                     /label=Q2
     primer_bind     complement(1178..1195)
                     /label=oBFC1708
     primer_bind     complement(1179..1213)
                     /label=oBFC2046
     primer_bind     complement(1185..1231)
                     /label=oBA1681_oBA1120_GG_pBA368_Gib_F
     primer_bind     complement(1191..1231)
                     /label=oBA1833_taCas13_specialGGBB_R
     primer_bind     complement(1192..1235)
                     /label=oBFC1698
     primer_bind     complement(1192..1235)
                     /label=PTet_R
     primer_bind     complement(1192..1235)
                     /label=oBA2200_pBA1169_BB_Gib_R
     primer_bind     complement(1192..1213)
                     /label=oBA1595_Cas13_BB_CommonGib_R
     primer_bind     complement(1192..1213)
                     /label=oBA1596_Cas13_BB_CommonGib_R
     primer_bind     complement(1194..1236)
                     /label=oBA2304_pTet_PCR_Gib_prep_R
     primer_bind     1204..1256
                     /label=oBA2343_ECOR31_Hachiman_Gib_p1_F
     RBS             1214..1231
                     /label=RBS
     misc_feature    1232..1233
                     /label=2>3
     misc_feature    1234..2017
                     /label=ECOR31_Hachiman
     CDS             1234..2013
                     /codon_start=1
                     /label=HamA (SAVED?)
                     /translation="MANFEDWCDSTERNISDHYLQSITARDAECMFGVQVMAALIPEHY
                     ASPRNIANAFEALGKPGLAAYIAGKLPETKQIRSGDLGEIFATEWINARSNGYKTPIKR
                     LRWKDHRNMSMRGEDVIGIYIDQSSQQLFFLKTEAKSRAKMTGEVVSEARDNLNKEQGL
                     PSSHALMFIADRLNEQGEELLAKAILNATLRQGIVPGCVRHLIFLLSGNSSETMLTTSI
                     EKYTGQNNQWGVCLRIARHGEFIAATFEKVISDASNS"
     primer_bind     complement(1995..2029)
                     /label=oBA2489_Ham31DELhamB_Mut_Gib_R
     CDS             1997..2017
                     /codon_start=1
                     /label=HamB
                     /translation="MPATAD"
     primer_bind     2000..2036
                     /label=oBA2488_Ham31DELhamB_Mut_Gib_F
     primer_bind     2017..2057
                     /label=oBA1713_p15_cmR_part4_gib_F
     primer_bind     2017..2036
                     /label=oBA1739_pt4_commonGib_F
     primer_bind     2017..2036
                     /label=oBA2199_pBA1169_BB_Gib_F
     primer_bind     2017..2033
                     /label=oBA1650_pBFC0924_Gib_F
     misc_feature    join(2018..3757,1..66)
                     /label=PCR from pBFC0335 or similar
     misc_feature    2018..2036
                     /label=Pt4_common_region_6>4
     misc_feature    2018..2036
                     /label=introduced
     primer_bind     2018..2036
                     /label=oBA2305_oBA1904_pt4ONLY_common_F
     misc_feature    2018..2021
                     /label=6>4 OH
     misc_feature    complement(2022..2025)
                     /label=SMRT Gate OH Right
     misc_feature    complement(2030..2036)
                     /label=AarI
     primer_bind     2036..2056
                     /label=oBFC1058_fixed
     primer_bind     2036..2056
                     /label=oBFC1060_fixed
     primer_bind     2036..2056
                     /label=oBFC1062_fixed
     primer_bind     2036..2056
                     /label=oBFC1447
     misc_feature    complement(2037..2145)
                     /label=CamR Terminator
     misc_feature    complement(2037..2145)
                     /label=CamR Terminator
     primer_bind     2037..2062
                     /label=oBA2031_WYL_p15a_CmR_Gib_F
     CDS             complement(2146..2805)
                     /label=CamR
     misc_feature    2572..2575
                     /label=BbsI OH
     promoter        complement(2806..2910)
                     /label=CamR Promoter
     promoter        complement(2806..2910)
                     /label=CamR Promoter
     rep_origin      complement(3178..3723)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     rep_origin      complement(3178..3723)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
ORIGIN
        1 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
       61 agttgtcacc tgcccgaacc agagacttat cctgaactgg ccgataattg cagacgaacg
      121 atgcttgggc ccgaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat
      181 catcatcatc atcattgagt ttaaacgctc tccagcttgg gtgttttggc ggatgagaga
      241 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
      301 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
      361 gccgtagcgc cgatggtagt gtggggtctg cccatgcgag agtaggcaac tgccaggcat
      421 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
      481 gtgaacttca gcgacgtctt aagacccact ttcacattta agttgttttt ctaatccgca
      541 tatgatcaat tcaaggccga ataagaaggc tggctctgca ccttggtgat caaataattc
      601 gatagcttgt cgtaataatg gcggcatact atcagtagta ggtgtttccc tttcttcttt
      661 agcgacttga tgctcttgat cttccaatac gcaacctaaa gtaaaatgcc ccacagcgct
      721 gagtgcatat aatgcattct ctagtgaaaa accttgttgg cataaaaagg ctaattgatt
      781 ttcgagagtt tcatactgtt tttctgtagg ccgtgtacct aaatgtactt ttgctccatc
      841 gcgatgactt agtaaagcac atctaaaact tttagcgtta ttacgtaaaa aatcttgcca
      901 gctttcccct tctaaagggc aaaagtgagt atggtgccta tctaacatct caatggctaa
      961 ggcgtcgagc aaagcccgct tattttttac atgccaatac aatgtaggct gctctacacc
     1021 tagcttctgg gcgagtttac gggttgttaa accttcgatt ccgacctcat taagcagctc
     1081 taatgcgctg ttaatcactt tacttttatc taatctagac atcattaatt cctaattttt
     1141 gttgacactc tatcgttgat agagttattt taccactccc tatcagtgat agagaaaaga
     1201 attcaaaaga tcttttaaga aggagatata ctgatggcaa atttcgagga ctggtgtgat
     1261 tcgacagagc gtaatatcag cgaccattat ttgcagtcaa taacggccag ggatgccgag
     1321 tgtatgttcg gcgttcaggt tatggccgcg ctaatacctg aacactatgc gtcaccaaga
     1381 aatatcgcta acgcatttga agctctaggt aaaccggggc tggcggcata cattgctgga
     1441 aaattgcctg aaacgaaaca gattcgttcc ggtgatttgg gtgagatatt tgcaactgaa
     1501 tggataaatg cacgtagcaa tggctataaa acgcctataa aaagattgcg ctggaaagat
     1561 catcgcaata tgtcgatgcg tggtgaggac gtcattggta tttacataga tcagtcttct
     1621 caacaacttt ttttccttaa aaccgaagcg aaaagtcgcg ccaaaatgac gggagaggtg
     1681 gtttctgaag cccgcgataa cttgaataaa gagcaagggc tgccatcttc acatgcccta
     1741 atgttcatag cagatcgcct gaatgaacag ggtgaggagc ttttggctaa agcgatctta
     1801 aatgcaacgc ttcggcaagg aattgttcca ggatgcgtcc ggcatcttat ctttctatta
     1861 tcgggcaact cttcagaaac tatgctgacc acttcaattg aaaagtatac aggtcaaaat
     1921 aatcagtggg gtgtttgtct aagaattgct cgccatggtg agtttattgc cgcaactttt
     1981 gaaaaggtga ttagcgatgc cagcaacagc tgattgaagg aggactttgg caggtgacca
     2041 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     2101 attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg
     2161 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     2221 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     2281 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     2341 actggtgaaa ctcacccagg gattggctga aacgaaaaac atattctcaa taaacccttt
     2401 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     2461 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     2521 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     2581 catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     2641 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     2701 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     2761 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     2821 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     2881 aaagttggaa cctcttacgt gcccgatcaa gcgctagcgg agtgtatact ggcttactat
     2941 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     3001 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     3061 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     3121 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     3181 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     3241 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     3301 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     3361 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     3421 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3481 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     3541 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     3601 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     3661 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     3721 caagaagatc atcttattaa tcagataaaa tatttct
//
