LOCUS       Exported                6517 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 6517)
  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..6517
                     /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..1253)
                     /label=oBA2487_Ham31DELhamA_Mut_Gib_R
     primer_bind     complement(1194..1236)
                     /label=oBA2304_pTet_PCR_Gib_prep_R
     RBS             1214..1231
                     /label=RBS
     primer_bind     1224..1263
                     /label=oBA2486_Ham31DELhamA_Mut_Gib_F
     misc_feature    1232..1233
                     /label=2>3
     misc_feature    1234..4777
                     /label=ECOR31_Hachiman
     CDS             1234..1269
                     /codon_start=1
                     /label=HamA (SAVED?)
                     /translation="MEKVISDASNS"
     CDS             1253..4777
                     /codon_start=1
                     /label=HamB
                     /translation="MPATADEIIEAIKEASAVGFRGRLIARGQARSVIWRDGDLPPDAP
                     EFSALLSQDLQGYAYALIDLGLRLRELNGDDAYARIAFEQAGTALESAIAKGKRDSRDT
                     DFHFVMAAASYHLAHLSARAYSLLAMVGQDDNFSPIERALTQLIRRDLRTLRDNALGFR
                     LRGDGSDVKITEILQARLNLPQDENGDSESEEDILFDGLDLALTDAYMSAISLYLLAVE
                     RGESRLLSRAIEKLRISLSICAQFNMLPQWWLNFITIHLLSDLWSDTFHERLPLVPVGG
                     DAAEWPALRELFIALLQRRPRAEIDLWPSQREAAGRSVNDNDDLVVSLPTSAGKTRIAE
                     LCILRCLAGGKRVVFITPLRALSAQTEATLSRTFGPLGKTISMLYGSIGVSGMDEDAIR
                     QRDIVVATPEKLDFALRNDPSIINDVGLFIFDEGHMIGADEREVRYEVQIQRLLRRQDA
                     DTRRIVCLSAILPDGEQLDDFAGWLRRDKPGGPIKNNWRPTRLQFGEVIWSAPAGRLNL
                     SVGYEAAWVSRFIVSRQPPKVKLPNKKQRTKMFPSDNKELCLATAWRLIEDGQTVLIYC
                     PLRRSVEPFAETIVDLHQRGLLPSLFDAAPDILDTAISLGEEWLGAHSPILACLRLGVA
                     LHHGALPTAYRKEIERLLRDGVLKVTISSPTLAQGLNLSATAIVMHSLHRNRELIKVSE
                     FRNVIGRAGRAYVDVEGLVIYPIFDKVNKRQTNWHTLTSDTGAREMESGLIQLVCVLLI
                     RMHTRLGGDLKALTEYVTNNAVAWEFPEIMTESPQERDIAQAIWEKQLSTLDTAILSLL
                     GENDIPDDQIETALDDILQSSLWQRSLQRYRDENERILLKSGLLSRSRYIWQRSTAAGR
                     RGYFLSGVGLTTGLRLDAIAAKANQLLIDANAAIMGGDAEEAIAAITALAEEVFTFYPF
                     IPDPLPGDWRGILRSWLLGEPMTNVANTQASETLQFVENGLVYRLPWAMEAIRVRATAN
                     GDLIGDTDTTLDDYELGFAVAAVETGTLSRSSSLLIQAGFSSRLAAIKVVTDTTADFQS
                     GQELRRWLNSEEVISHTDNHDWPTPETRVMWLEFLGSLSPKGSQVWSRHRYNGMVDWRD
                     TPAVIGTPLQLYTVDGIHHVLADDGTPLGSINGRINTNRRGLLRVEVDDENGRAMFDYL
                     GPDDFIST"
     primer_bind     complement(2859..2892)
                     /label=oBA2344_ECOR31_Hachiman_Gib_p1_R
     primer_bind     2859..2892
                     /label=oBA2345_ECOR31_Hachiman_Gib_p2_F
     primer_bind     complement(4752..4807)
                     /label=oBA2346_ECOR31_Hachiman_Gib_p2_R
     primer_bind     4777..4817
                     /label=oBA1713_p15_cmR_part4_gib_F
     primer_bind     4777..4796
                     /label=oBA1739_pt4_commonGib_F
     primer_bind     4777..4796
                     /label=oBA2199_pBA1169_BB_Gib_F
     primer_bind     4777..4793
                     /label=oBA1650_pBFC0924_Gib_F
     misc_feature    join(4778..6517,1..66)
                     /label=PCR from pBFC0335 or similar
     misc_feature    4778..4796
                     /label=Pt4_common_region_6>4
     misc_feature    4778..4796
                     /label=introduced
     primer_bind     4778..4796
                     /label=oBA2305_oBA1904_pt4ONLY_common_F
     misc_feature    4778..4781
                     /label=6>4 OH
     misc_feature    complement(4782..4785)
                     /label=SMRT Gate OH Right
     misc_feature    complement(4790..4796)
                     /label=AarI
     primer_bind     4796..4816
                     /label=oBFC1058_fixed
     primer_bind     4796..4816
                     /label=oBFC1060_fixed
     primer_bind     4796..4816
                     /label=oBFC1062_fixed
     primer_bind     4796..4816
                     /label=oBFC1447
     misc_feature    complement(4797..4905)
                     /label=CamR Terminator
     misc_feature    complement(4797..4905)
                     /label=CamR Terminator
     primer_bind     4797..4822
                     /label=oBA2031_WYL_p15a_CmR_Gib_F
     CDS             complement(4906..5565)
                     /label=CamR
     misc_feature    5332..5335
                     /label=BbsI OH
     promoter        complement(5566..5670)
                     /label=CamR Promoter
     promoter        complement(5566..5670)
                     /label=CamR Promoter
     rep_origin      complement(5938..6483)
                     /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(5938..6483)
                     /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 ctgatggaaa aggtgattag cgatgccagc
     1261 aacagctgat gaaattatcg aagctattaa ggaagcttca gctgtaggtt ttcgcgggcg
     1321 cctgattgca cgcgggcaag cccggtcggt gatctggcga gacggtgact tgccgccgga
     1381 tgcaccggag tttagcgcac tgctcagtca ggaccttcaa ggctatgctt atgccctaat
     1441 cgatctcggc cttcgtcttc gagaacttaa tggagacgac gcctatgcgc gaattgcttt
     1501 tgaacaggcc ggaaccgcac tcgagtcggc tattgctaaa ggaaaacgcg acagccggga
     1561 tacagatttt cacttcgtaa tggctgctgc aagctatcac cttgcgcatc tttcagcacg
     1621 agcctattcg ttactcgcaa tggttggtca ggatgataat ttttcaccaa tagagagggc
     1681 tcttacacaa ctgatccggc gtgatttgcg tactctgcgc gataatgcgt tgggctttcg
     1741 gctcagaggt gatggcagtg acgtgaaaat tactgaaatc ctccaggctc gcctaaactt
     1801 gccgcaagac gagaacggtg acagtgagtc agaggaggac atcctttttg atgggcttga
     1861 tttggcactg actgacgcct atatgtcggc catttctctc taccttctcg ctgtagaaag
     1921 gggagaatca cgtttgcttt ccagggctat tgaaaaatta agaattagtc taagtatatg
     1981 tgctcagttt aatatgctac cccagtggtg gctaaacttt attaccattc atctgttatc
     2041 tgacctttgg tcagatacat tccatgaaag attgccactg gttccagttg gtggtgacgc
     2101 tgctgaatgg cccgcgcttc gggaactgtt tattgcactg cttcaaagac gacctcgagc
     2161 cgaaattgat ctctggccat cgcagagaga ggccgccgga cgctctgtta atgacaatga
     2221 cgatttggtt gtttcgttgc caaccagcgc tggaaaaact cgtattgctg aattgtgtat
     2281 tcttcgctgt ctggcgggtg gcaagcgtgt tgttttcatc accccgcttc gtgcgctttc
     2341 tgcacaaaca gaagccactc tctccagaac atttgggccg cttgggaaaa caatatcaat
     2401 gctgtatggc agcattggcg taagtggtat ggatgaggat gcaattcgcc agcgtgacat
     2461 cgtcgtcgcc acgcctgaaa aacttgactt tgctctccgg aatgacccgt caattattaa
     2521 tgatgtagga ctttttattt ttgatgaggg gcacatgatt ggggccgatg aacgagaggt
     2581 tcggtatgaa gttcaaattc aacggctgtt gcgcagacag gatgccgata cgcgcagaat
     2641 tgtctgcctc tcggcaattc tacctgatgg tgagcaactc gatgattttg ccggttggct
     2701 ccgtcgtgac aagcccggag gaccaatcaa aaataattgg cgaccgaccc gcttgcagtt
     2761 tggcgaggtc atatggtccg ctccagcagg tcggcttaat ctcagcgtcg gttatgaggc
     2821 ggcttgggta tcgcgtttta tcgtcagcag acaaccgcca aaggttaaat tacctaataa
     2881 aaagcagcga acaaaaatgt ttccctctga taacaaggag ctctgcttag cgacagcctg
     2941 gcgtctaatt gaagatggac agaccgtact tatctactgc ccacttcgcc ggagcgtcga
     3001 accgtttgcc gagacaattg tcgatctgca tcagcgcgga ctactgccat ctttatttga
     3061 tgctgccccg gacattctgg atacggctat ctcgctggga gaagaatggt tgggggcaca
     3121 cagcccgata cttgcctgcc tacggttggg cgtggctctt catcatggcg cattgcctac
     3181 ggcttaccgc aaagaaatcg aacgtttgct gcgagatggg gtacttaaag tgacgatctc
     3241 atcaccgacg ctagcacaag gtcttaatct ttcggcaaca gctatcgtca tgcattctct
     3301 tcatcggaat cgcgagctta tcaaagtgtc cgagttcaga aacgtcatag gcagagcagg
     3361 gcgcgcatat gtagatgttg aagggctggt gatttatcca atattcgata aagtaaataa
     3421 acgccagact aactggcata ccctcacatc tgacacgggg gcccgtgaga tggagagcgg
     3481 actaatacag cttgtctgtg tgttattgat tcgcatgcat acacggcttg gtggggattt
     3541 aaaagcactc actgaatatg tgaccaacaa cgccgtagcg tgggaatttc cagaaattat
     3601 gaccgaatct ccgcaagaaa gagatattgc gcaggccatc tgggaaaaac aactttcaac
     3661 gcttgatact gctattctga gccttcttgg tgagaatgat atcccggatg accagattga
     3721 aactgctctt gatgacattc ttcagtcttc actttggcaa cgcagtttgc agcgttatag
     3781 ggatgagaat gaacggattt tgctgaaatc gggactcctg tcccgcagcc gctacatctg
     3841 gcagcgctca acggcagcag ggcgccgggg gtatttcctg tcgggggtag gacttacaac
     3901 cgggctacga cttgatgcta ttgccgctaa ggccaaccaa ctattgattg atgcgaatgc
     3961 ggctataatg ggcggtgatg ccgaagaggc aattgctgct atcacggcgc tcgctgaaga
     4021 ggtctttacc ttctatccgt ttatacccga tccattgcct ggcgactgga gaggtatttt
     4081 acgttcctgg ctattgggtg aacccatgac taacgtcgcg aatacgcagg cgtctgaaac
     4141 gcttcaattc gtcgagaatg ggctggtgta tcgacttccg tgggcaatgg aggctattag
     4201 ggtacgtgca acagctaatg gagacctcat aggtgataca gataccacgc tagacgatta
     4261 tgagttaggt tttgcggttg ctgctgttga aacaggtacc ttgagccgtt cgtcttcact
     4321 gcttattcag gcgggtttta gctctaggct tgcagccata aaagtggtta cagacactac
     4381 tgccgatttt caatccggcc aggagcttag aaggtggctg aactctgaag aagtgataag
     4441 ccacacggat aatcatgact ggcctacacc tgaaaccaga gtgatgtggc tggaattctt
     4501 gggtagtctc agccccaaag gcagccaggt ttggagtcgt catcgttata acggaatggt
     4561 ggactggaga gatactccgg cagtgatcgg cacaccattg cagctttaca ctgttgatgg
     4621 tattcatcac gtactggctg atgatggcac tcctctggga agtataaatg gcagaattaa
     4681 cacaaacaga agaggtcttc ttagagttga ggtggatgat gagaatggtc gtgccatgtt
     4741 cgattattta ggaccggacg actttatttc tacctgaagg aggactttgg caggtgacca
     4801 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     4861 attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg
     4921 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     4981 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     5041 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     5101 actggtgaaa ctcacccagg gattggctga aacgaaaaac atattctcaa taaacccttt
     5161 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     5221 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     5281 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     5341 catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     5401 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     5461 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     5521 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     5581 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     5641 aaagttggaa cctcttacgt gcccgatcaa gcgctagcgg agtgtatact ggcttactat
     5701 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     5761 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     5821 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     5881 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     5941 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     6001 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     6061 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     6121 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     6181 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6241 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     6301 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     6361 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     6421 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     6481 caagaagatc atcttattaa tcagataaaa tatttct
//
