LOCUS       Exported                6244 bp ds-DNA     circular SYN 26-NOV-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6244)
  AUTHORS   Doudna lab
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, May 10, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6244
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    complement(139..167)
                     /label=AmpR_promoter
     misc_feature    209..1069
                     /label=AMP resistance beta-lactimase
     CDS             407..1066
                     /label=AmpR
     primer_bind     827..848
                     /label=GJK_331
     primer_bind     complement(851..872)
                     /label=GJK_332
     rep_origin      1164..1846
                     /label=ColE1 origin
     misc_feature    complement(1276..1385)
                     /label=RNA1-for
     misc_feature    complement(1824..1833)
                     /label=ColE1 ORI
     primer_bind     2043..2060
                     /label=GJK_388
     primer_bind     complement(2061..2081)
                     /label=GJK_399
     misc_feature    complement(2257..2482)
                     /label=ROP/ROM
     primer_bind     4010..4029
                     /label=T7
     CDS             4147..5349
                     /codon_start=1
                     /label=MBP ORF
                     /translation="MKSSHHHHHHGSSMKIEEGKLVIWINGDKGYNGLAEVGKKFEKDT
                     GIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLY
                     PFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMF
                     NLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADT
                     DYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGIN
                     AASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQK
                     GEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGIEE
                     NL"
     misc_feature    4159..4176
                     /label=His6 tag
     primer_bind     complement(5339..5387)
                     /label=JW44
     primer_bind     complement(5341..5371)
                     /label=TEV_GABPA_R
                     /note="Reverse primer to amplify 2BT vector from TEV site
                     Overlapping region complementary to N-terminus of GABPA 
                     gBlock"
     primer_bind     complement(5341..5371)
                     /label=TEV_GABPB1L_R
                     /note="Reverse primer to amplify 2BT vector from TEV site
                     Overlapping region complementary to N-terminus of GABPB"
     primer_bind     complement(5341..5364)
                     /label=GJK_387
     primer_bind     5348..5396
                     /label=JW45
     misc_feature    5350..5364
                     /label=Lic_v1_For
     CDS             5368..5658
                     /codon_start=1
                     /product="Tax=CG_Thiomicrospira_01"
                     /label=Cas2
                     /db_xref="mme:Marme_0668"
                     /translation="MKHYLICFDVQHDKTRAKLSRLLEKYGPRVQGSVFEVSFKTPDRK
                     RQLEYKIHQIIKQSNTEENNIRFYNLNKDTIKHSHDINGNPIAQLPAAIVL"
     gene            5368..5658
                     /locus_tag="CG02_land_8_20_14_3.00_150_scaffold_47_76"
                     /label=CG02_land_8_20_14_3.00_150_scaffold_47_76
     CDS             5368..5371
                     /codon_start=1
                     /product="Tax=CG_Thiomicrospira_01"
                     /label=RT-Cas1
                     /note="/modified_by=User"
                     /db_xref="33602764"
                     /db_xref="mme:Marme_0669"
                     /db_xref="UniRef100_F2K1V9"
                     /db_xref="K15342"
                     /translation=""
     gene            5368..5371
                     /locus_tag="CG02_land_8_20_14_3.00_150_scaffold_47_75"
                     /label=CG02_land_8_20_14_3.00_150_scaffold_47_75
     primer_bind     complement(5630..5678)
                     /label=JW46
     primer_bind     5633..5681
                     /label=JW43
     primer_bind     5659..5682
                     /label=GJK_386
     misc_feature    5659..5673
                     /label=Lic_v1_rev
     primer_bind     5662..5682
                     /label=Termination_GABPB1L_F
                     /note="Forward primer to amplify 2BT vector at translation 
                     termination
                     Overlapping region complementary to C-terminus of GABPB"
     primer_bind     5662..5682
                     /label=Termination_GABPA_F
                     /note="Forward primer to amplify 2BT vector at translation 
                     terminationOverlapping region complementary to C-terminus 
                     of GABPA"
     misc_feature    5780..5798
                     /label=T7 terminator primer
     misc_feature    5801..5841
                     /label=T7 terminator
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa cattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
     2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
     3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
     3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
     3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt
     3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt
     3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg
     3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
     3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc
     3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gacgctctcc
     3721 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     3781 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     3841 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     3901 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     3961 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     4021 actataggga gaccacaacg gtttccctct agtgccggct ccggagagct ctttaattaa
     4081 gcggccgccc tgcaggactc gagttctaga aataattttg tttaacttta agaaggagat
     4141 atacatatga aatcttctca ccatcaccat caccatggtt cttctatgaa aatcgaagaa
     4201 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt
     4261 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa
     4321 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac
     4381 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg
     4441 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt
     4501 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac
     4561 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag
     4621 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac
     4681 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat
     4741 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg
     4801 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg
     4861 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta
     4921 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca
     4981 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg
     5041 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg
     5101 aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc
     5161 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt
     5221 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg
     5281 cagactaatt cgagctcgaa caacaacaac aataacaata acaacaacct cgggatcgag
     5341 gaaaacctgt acttccaatc caatgctatg aaacactacc tgatttgctt tgatgtccaa
     5401 cacgacaaaa cccgcgcaaa actctcaagg ctgctggaaa aatacggccc cagagtccaa
     5461 ggctcggtat ttgaagtcag tttcaaaacc cccgatcgca aacgacaact agaatacaaa
     5521 atccaccaaa ttatcaaaca atccaacacc gaagaaaaca atatccgttt ctacaacctc
     5581 aataaagaca ccataaaaca cagtcacgac atcaatggca atcccatcgc tcaattacct
     5641 gctgcaatcg ttctataaat tggaagtgga taacggatcc gcgatcgcgg cgcgccacct
     5701 ggtggccggc cggtaccacg cgtgcgcgct gatccggctg ctaacaaagc ccgaaaggaa
     5761 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     5821 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc acaggacggg
     5881 tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga gcaggactgg
     5941 gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa attgcatcaa
     6001 cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat ggacgatatc
     6061 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac
     6121 ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta
     6181 gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg
     6241 agaa
//
