LOCUS       pYZ038_Topo_clon        3673 bp DNA     circular SYN 15-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3673)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 15, 2023 from SnapGene 6.2.2
            https://www.snapgene.com
COMMENT     pCR blunt II TOPO from 1 to 3519
            >pCR blunt II TOPO
FEATURES             Location/Qualifiers
     source          1..3673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(908..1282)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             complement(1489..2283)
                     /label=Kan/neoR
     promoter        2284..2421
                     /label=kan promoter
     CDS             complement(2632..2934)
                     /label=ccdB ORF
     misc_feature    2935..2943
                     /label=fusion linker
     primer_bind     3072..3088
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3095..3113
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        3185..3260
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3185..3209
                     /label=annealing
     misc_feature    3261..3266
                     /label=linker
     misc_feature    3267..3337
                     /label=tRNA-Gly
     misc_feature    complement(3313..3337)
                     /label=annealing
     CDS             complement(3338..3457)
                     /label=LacZ alpha ORF
     promoter        complement(3417..3435)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3453..3469)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3458..3579
                     /label=lac promoter/operator promoter
                     /label=lac promoter/operator
     protein_bind    3477..3493
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3501..3531)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3546..3567
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaagccca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 gcacgtgtca gtcctgctcc tcggccacga agtgcacgca gttgccggcc gggtcgcgca
      961 gggcgaactc ccgcccccac ggctgctcgc cgatctcggt catggccggc ccggaggcgt
     1021 cccggaagtt cgtggacacg acctccgacc actcggcgta cagctcgtcc aggccgcgca
     1081 cccacaccca ggccagggtg ttgtccggca ccacctggtc ctggaccgcg ctgatgaaca
     1141 gggtcacgtc gtcccggacc acaccggcga agtcgtcctc cacgaagtcc cgggagaacc
     1201 cgagccggtc ggtccagaac tcgaccgctc cggcgacgtc gcgcgcggtg agcaccggaa
     1261 cggcactggt caacttggcc atggtggccc tcctcacgtg ctattattga agcatttatc
     1321 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1381 gggttccgcg cacatttccc cgaaaagtgc cacctgtatg cggtgtgaaa taccgcacag
     1441 atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaataattc agaagaactc
     1501 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     1561 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     1621 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     1681 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     1741 gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     1801 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     1861 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     1921 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     1981 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     2041 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     2101 ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     2161 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     2221 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     2281 catgcgaaac gatcctcatc ctgtctcttg atcagagctt gatcccctgc gccatcagat
     2341 ccttggcggc gagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg
     2401 cgccccagct ggcaattccg gttcgcttgc tgtccataaa accgcccagt ctagctatcg
     2461 ccatgtaagc ccactgcaag ctacctgctt tctctttgcg cttgcgtttt cccttgtcca
     2521 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     2581 cgtgaaaagg atctaggtga agatcctttt tgataatctc atgcctgaca tttatattcc
     2641 ccagaacatc aggttaatgg cgtttttgat gtcattttcg cggtggctga gatcagccac
     2701 ttcttccccg ataacggaga ccggcacact ggccatatcg gtggtcatca tgcgccagct
     2761 ttcatccccg atatgcacca ccgggtaaag ttcacgggag actttatctg acagcagacg
     2821 tgcactggcc agggggatca ccatccgtcg ccccggcgtg tcaataatat cactctgtac
     2881 atccacaaac agacgataac ggctctctct tttataggtg taaaccttaa actgccgtac
     2941 gtataggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     3001 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     3061 gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat agggcgaatt
     3121 gggccctcta gatgcatgct cgagcggccg ccagtgtgat ggatatctgc agaattcgcc
     3181 cttagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
     3241 aagtggcacc gagtcggtgc aaacaagcgc aagtggttta gtggtaaaat ccaacgttgc
     3301 catcgttggg cccccggttc gattccgggc ttgcgcaaag ggcgaattcc agcacactgg
     3361 cggccgttac tagtggatcc gagctcggta ccaagcttga tgcatagctt gagtattcta
     3421 tagtgtcacc taaatagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3481 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3541 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3601 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3661 cgtattgggc gct
//
