LOCUS       pC1.530_Curing_v        6420 bp DNA     circular SYN 10-NOV-2023
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6420)
  AUTHORS   SynthSys Center
  TITLE     Direct Submission
  JOURNAL   Exported Nov 10, 2023 from SnapGene 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6420
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     misc_feature    34..533
                     /label=UP FLANK
                     /note="pUC 19 will go here"
     misc_feature    534..537
                     /label=AATG
     misc_feature    538..572
                     /label=Bba_J23119
                     /note="'>'>http://parts.igem.org/Part:BBa_J23119"
     misc_feature    561..566
                     /label=-10
     misc_feature    573
                     /label=TSS start site
                     /note="'>'>http://parts.igem.org/Part:BBa_K1824556"
     misc_feature    574..589
                     /label=leader sequence
                     /label=nonstandard type: leader
     misc_feature    590..593
                     /label=Junction 1
     misc_feature    594..612
                     /label=DR
     misc_feature    613..634
                     /label=Cas12a gRNA
     misc_feature    639..656
                     /label=crRNA2
     misc_feature    639..656
                     /label=Direct repeat
                     /label=nonstandard type: DR
     misc_feature    657
                     /label=Spacer 4
                     /label=nonstandard type: Spacer
     misc_feature    658..718
                     /label=Terminator (L3S2P21)
     misc_feature    719..722
                     /label=AGGT
     misc_feature    723..1573
                     /label=GmR cassette
     misc_feature    complement(1576..1577)
                     /label=Promoter
     misc_feature    1578..2077
                     /label=DOWN Flank
     misc_feature    complement(2109..2236)
                     /label=RB short
                     /label=RB\short
                     /note="vntifkey=21"
     misc_feature    complement(2260..3741)
                     /label=RK2 trfa (no Esp3I)
                     /label=RK2\trfa\(no\Esp3I)
                     /note="vntifkey=21"
     misc_feature    3752..4541
                     /label=pUC ori
                     /label=pUC\ori
                     /note="vntifkey=21"
     rep_origin      3787
                     /label=ORI
                     /note="RNaseH cleavage point
                     vntifkey=33"
     CDS             complement(4549..5406)
                     /codon_start=1
                     /label=AP r
                     /label=AP\r
                     /note="vntifkey=4"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(5604..6221)
                     /direction=LEFT
                     /standard_name="oriV"
                     /label=oriV
                     /note="shows similarity to GenBank Accession Number M20134 
                     vntifkey=33"
     misc_feature    6251..6398
                     /label=TDNA-LB
                     /note="shows similarity to T-DNA left border: GenBank
                     Accession Number J01825
                     vntifkey=21"
ORIGIN
        1 tctgtgaaga caatgccgaa ttcggatccg gagtgaccat ctcatctgta acatcattgg
       61 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
      121 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
      181 cagcatccat gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc
      241 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
      301 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa
      361 taaatcgaac ttttgctgag ttgaaggatc agctcgagtt accaatgctt aatcagtgag
      421 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
      481 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataatgttg
      541 acagctagct cagtcctagg tataatgcta gcagctgatt taggcaaaaa cggaatttct
      601 actgttgtag atttaataaa tatagtttaa gtaaaacgaa tttctactgt tgtagatctc
      661 ggtaccaaat tccagaaaag aggcctcccg aaaggggggc cttttttcgt tttggtccag
      721 gtggtcgctt tcaggggtac cgagctcgaa ttggccgcgg cgttgtgaca atttaccgaa
      781 caactccgcg gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg
      841 gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga gagccactgc
      901 gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg cgttggcctc
      961 atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct cgccggagac
     1021 tgcgagatca tagatataga tctcactacg cggctgctca aacctgggca gaacgtaagc
     1081 cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga atgtcttact
     1141 acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg tggctacgtc
     1201 tccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt ggtcagggcc
     1261 gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt tagggcgact
     1321 gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt gctgctccat aacatcaaac
     1381 atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa
     1441 aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca
     1501 aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac
     1561 aggcttatgt caagctttac tggagttatt aacactgtgt acccggacaa acaccattaa
     1621 tcagcaggaa ggtaaattct tcaatatcct gttcaacggt caactgttcg gtcagcaggc
     1681 gataccaaca aaaaccaaaa atcatatcca gcagcagttc acgattgata tctttcggca
     1741 gttcaccatt gctaatggca tcttcaacca gttttttcgg tatctcacga cgacgttcca
     1801 taaactgatc tttcagttgg gtcagggtta cagggtccaa ctgtgcttct gcaataacac
     1861 aacgaaatgc ttcaccacaa atggtttcac gccaaacttt ccacagatta tgcagcagaa
     1921 aatccagatc ggctttaaag ctacccaaat ccggaaattt acgtacctgt tcgatttcat
     1981 tttcatacac ttcggcaatc agtgctgctt tgttggtcca ccaacgataa atggtcggtt
     2041 tgcctgcacc ggcgcgacgt gccacgcttt caatgctcgc tgcaattgtc ttctgcacga
     2101 agtggtttaa actatcagtg tttgacagga tatattggcg ggtaaaccta agagaaaaga
     2161 gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat
     2221 ttgtatgtgc atgccaacca cagggttccc cagatcaggc gctggctgct gaacccccag
     2281 ccggaactga ccccacaagg ccctagcgtt tgcaatgcac caggtcatca ttgacccagg
     2341 cgtgttccac caggccgctg cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc
     2401 acttcttcac gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc agcttgagcg
     2461 ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct
     2521 tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc acgacgattt
     2581 cctcgtcgat caggacctgg caacgggacg ttttcttgcc acggtccagg acgcggaagc
     2641 ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg
     2701 tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta ataccggcca ttgatcgacc
     2761 agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc
     2821 gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga
     2881 cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg
     2941 cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg
     3001 gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa caaggaaagc tgcatttcct
     3061 tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg acctgttttg
     3121 ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg
     3181 tcatcgactt cgccaaacct gccgcctcct gttcaagacg acgcgaacgc tccacggcgg
     3241 ccgatggcgc gggcagggca gggggagcca gttgcacgct gtcgcgctcg atcttggccg
     3301 tagcttgctg gaccatcgag ccgacggact ggaaggtttc gcggggcgca cgcatgacgg
     3361 tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc
     3421 tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt gccccgactc
     3481 acgccggggc aatgtgccct tattcctgat ttgacccgcc tggtgccttg gtgtccagat
     3541 aatccacctt atcggcaatg aagtcggtcc cgtagaccgt ctggccgtcc ttctcgtact
     3601 tggtattccg aatcttgccc tgcacgaata ccagcgaccc cttgcccaaa tacttgccgt
     3661 gggcctcggc ctgagagcca aaacacttga tgcggaagaa gtcggtgcgc tcctgcttgt
     3721 cgccggcatc gttgcgccac atctaggatc tgccaggaac cgtaaaaagg ccgcgttgct
     3781 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3841 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3901 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3961 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4021 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4081 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4141 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4201 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4261 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4321 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4381 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4441 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4501 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4561 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4621 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4681 accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4741 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4801 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4861 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4921 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4981 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5041 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5101 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5161 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5221 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5281 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5341 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5401 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5461 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacgaattg
     5521 gccagcgctg ccatttttgg ggtgaggccg ttcgcggccg aggggcgcag cccctggggg
     5581 gatgggaggc ccgcgttagc gggccgggag ggttcgagaa gggggggcac cccccttcgg
     5641 cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta taaatattgg
     5701 tttaaaagca ggttaaaaga caggttagcg gtggccgaaa aacgggcgga aacccttgca
     5761 aatgctggat tttctgcctg tggacagccc ctcaaatgtc aataggtgcg cccctcatct
     5821 gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag tagtcgcgcc
     5881 cctcaagtgt caataccgca gggcacttat ccccaggctt gtccacatca tctgtgggaa
     5941 actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc acgtcgccgg
     6001 ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg cccctcaagt
     6061 gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt atccacaacg
     6121 ccggcggccg cggtgtctcg cacacggctt cgacggcgtt tctggcgcgt ttgcagggcc
     6181 atagacggcc gccagcccag cggcgagggc aaccagcccg gtgagcgtcg caaaggagat
     6241 cctgatctga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg
     6301 gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacaaatt gacgcttaga
     6361 caacttaata acacattgcg gacgttttta atgtactggg gtggatgcag tgggccccac
//
