LOCUS       pCHC005        7671 bp DNA     circular SYN 30-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7671)
  AUTHORS   CCALVEY
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2024 from SnapGene 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     -35_signal      134..139
                     /label=-35 kanR
     -10_signal      157..162
                     /label=-10
     primer_bind     complement(254..279)
                     /label=oCHC026: pK18sB seq R
                     /note="Sequencing  inserts in pK18sB MCS."
     CDS             266..1060
                     /gene="nptII"
                     /product="KanR (neomycin phosphotransferase)"
                     /label=KanR (neomycin phosphotransferase)
                     /label=neomycin\phosphotransferase
                     /note="/vntifkey=4"
                     /db_xref="GI:215434895"
                     /protein_id="ACJ66844.1"
     CDS             1552..2973
                     /gene="sacB"
                     /product="Levan Sucrase"
                     /label=SacB (levan sucrase)
                     /label=levan\sucrase
                     /note="/vntifkey=4"
                     /db_xref="GI:215434896"
                     /protein_id="ACJ66845.1"
     oriT            3374..3443
                     /label=RP4 oriT
                     /label=RP4\oriT
                     /note="RP4 oriT"
                     /note="/vntifkey=104"
     rep_origin      4329..5011
                     /label=ColE1 origin
     primer_bind     4914..4935
                     /label=oCHC025: pK18sB seq F
                     /note="Sequencing  inserts in pK18sB MCS."
     primer_bind     complement(5289..5330)
                     /label=oCHC098: pK18 mobSacB R
                     /note="Amplification  of pK18mobSacB vector backbone."
     gene            5331..6351
                     /locus_tag="H16_RS33875"
                     /old_locus_tag="H16_A0005"
                     /label=H16_RS33875
     misc_feature    5331..6351
                     /label=UP
                     /label=UP r1
                     /note="1kb H16_A0006 (Type I restriction-modification 
                     system restriction subunit) Upstream Targeting Region"
     primer_bind     5632..5651
                     /label=oCHC029: RE Up Seq
                     /note="Sequencing  of pCHC005 Upstream targeting region."
     primer_bind     complement(6329..6351)
                     /label=oCHC028: RE ko R
                     /note="PCR  Mutagenesis to reduce size of ORNL 
                     pK18mobsacB_H16_0006_polyattB."
     primer_bind     6352..6384
                     /label=oCHC027: RE ko F
                     /note="PCR  Mutagenesis to reduce size of ORNL 
                     pK18mobsacB_H16_0006_polyattB, adds Pme1  Site."
     misc_feature    6352..6359
                     /label=PmeI
     misc_feature    6360..7406
                     /label=DOWN
                     /label=DN
                     /note="1kb H16_A0006 (Type I restriction-modification 
                     system restriction subunit) Downstream Targeting Region"
     gene            6360..7072
                     /locus_tag="H16_RS00035"
                     /old_locus_tag="H16_A0007"
                     /label=H16_RS00035
     gene            7079..7406
                     /locus_tag="H16_RS00040"
                     /old_locus_tag="H16_A0008"
                     /label=H16_RS00040
     primer_bind     complement(7132..7153)
                     /label=oCHC030: RE Down Seq
                     /note="Sequencing of pCHC005 Downstream targeting region."
     primer_bind     7407..7432
                     /label=oCHC097: pK18 mobSacB F
                     /note="Amplification of pK18mobSacB vector backbone."
     CDS             7504..7572
                     /label=LacZ alpha
ORIGIN
        1 ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagagaaa
       61 gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt tatggacagc
      121 aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt
      181 aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gatctgatca
      241 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
      301 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
      361 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
      421 cctgtccggt gccctgaatg aactccaaga cgaggcagcg cggctatcgt ggctggccac
      481 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
      541 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
      601 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
      661 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
      721 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
      781 caggctcaag gcgcggatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
      841 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
      901 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
      961 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     1021 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgct
     1081 agaggatcga tcctttttaa cccatcacat atacctgccg ttcactatta tttagtgaaa
     1141 tgagatatta tgatattttc tgaattgtga ttaaaaaggc aactttatgc ccatgcaaca
     1201 gaaactataa aaaatacaga gaatgaaaag aaacagatag attttttagt tctttaggcc
     1261 cgtagtctgc aaatcctttt atgattttct atcaaacaaa agaggaaaat agaccagttg
     1321 caatccaaac gagagtctaa tagaatgagg tcgaaaagta aatcgcgcgg gtttgttact
     1381 gataaagcag gcaagaccta aaatgtgtaa agggcaaagt gtatactttg gcgtcacccc
     1441 ttacatattt taggtctttt tttattgtgc gtaactaact tgccatcttc aaacaggagg
     1501 gctggaagaa gcagaccgct aacacagtac ataaaaaagg agacatgaac gatgaacatc
     1561 aaaaagtttg caaaacaagc aacagtatta acctttacta ccgcactgct ggcaggaggc
     1621 gcaactcaag cgtttgcgaa agaaacgaac caaaagccat ataaggaaac atacggcatt
     1681 tcccatatta cacgccatga tatgctgcaa atccctgaac agcaaaaaaa tgaaaaatat
     1741 caagtttctg aatttgattc gtccacaatt aaaaatatct cttctgcaaa aggcctggac
     1801 gtttgggaca gctggccatt acaaaacgct gacggcactg tcgcaaacta tcacggctac
     1861 cacatcgtct ttgcattagc cggagatcct aaaaatgcgg atgacacatc gatttacatg
     1921 ttctatcaaa aagtcggcga aacttctatt gacagctgga aaaacgctgg ccgcgtcttt
     1981 aaagacagcg acaaattcga tgcaaatgat tctatcctaa aagaccaaac acaagaatgg
     2041 tcaggttcag ccacatttac atctgacgga aaaatccgtt tattctacac tgatttctcc
     2101 ggtaaacatt acggcaaaca aacactgaca actgcacaag ttaacgtatc agcatcagac
     2161 agctctttga acatcaacgg tgtagaggat tataaatcaa tctttgacgg tgacggaaaa
     2221 acgtatcaaa atgtacagca gttcatcgat gaaggcaact acagctcagg cgacaaccat
     2281 acgctgagag atcctcacta cgtagaagat aaaggccaca aatacttagt atttgaagca
     2341 aacactggaa ctgaagatgg ctaccaaggc gaagaatctt tatttaacaa agcatactat
     2401 ggcaaaagca catcattctt ccgtcaagaa agtcaaaaac ttctgcaaag cgataaaaaa
     2461 cgcacggctg agttagcaaa cggcgctctc ggtatgattg agctaaacga tgattacaca
     2521 ctgaaaaaag tgatgaaacc gctgattgca tctaacacag taacagatga aattgaacgc
     2581 gcgaacgtct ttaaaatgaa cggcaaatgg tacctgttca ctgactcccg cggatcaaaa
     2641 atgacgattg acggcattac gtctaacgat atttacatgc ttggttatgt ttctaattct
     2701 ttaactggcc catacaagcc gctgaacaaa actggccttg tgttaaaaat ggatcttgat
     2761 cctaacgatg taacctttac ttactcacac ttcgctgtac ctcaagcgaa aggaaacaat
     2821 gtcgtgatta caagctatat gacaaacaga ggattctacg cagacaaaca atcaacgttt
     2881 gcgccgagct tcctgctgaa catcaaaggc aagaaaacat ctgttgtcaa agacagcatc
     2941 cttgaacaag gacaattaac agttaacaaa taaaaacgca aaagaaaatg ccgatgggta
     3001 ccgagcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     3061 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccct cgcggacgtg
     3121 ctcatagtcc acgacgcccg tgattttgta gccctggccg acggccagca ggtaggccga
     3181 caggctcatg ccggccgccg ccgccttttc ctcaatcgct cttcgttcgt ctggaaggca
     3241 gtacaccttg ataggtgggc tgcccttcct ggttggcttg gtttcatcag ccatccgctt
     3301 gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag
     3361 caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac
     3421 ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct
     3481 ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat
     3541 gaccccgaag cagggttatg cagcggaaaa gcgctgcttc cctgctgttt tgtggaatat
     3601 ctaccgactg gaaacaggca aatgcaggaa attactgaac tgaggggaca ggcgagagac
     3661 gatgccaaag agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     3721 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa
     3781 agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt
     3841 gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac
     3901 tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctcc
     3961 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     4021 gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt
     4081 cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg
     4141 tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt agtgtatgat
     4201 ggtgtttttg aggtgctcca gtggcttctg tttctatcag ggctggatga tcctccagcg
     4261 cggggatctc atgctggagt tcttcgccca ccccaaaagg atctaggtga agatcctttt
     4321 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4381 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4441 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4501 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4561 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4621 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4681 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4741 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4801 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4861 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4921 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4981 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5041 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5101 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5161 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5221 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     5281 taatgtgagt tagctcactc aggaaacagc tatgacatga ttacgaattc gacgatgacg
     5341 aagatttctc cgagcagtgt ttttcgtccg gcatactttt ttcatgtcgc caagcaagct
     5401 gagcgattcc tgaaggctca aaccaatggc actggcattc cacacgttga ccgagagctt
     5461 ctcgagggga taaaggtctt ttgtcctggc tctacggagc agcaattact tgcggaaatc
     5521 ctcgacactc tcgacaccgc catctacgaa actgaagcga tcatcgccaa gctcaaggcg
     5581 gtcaagcaag gcctgctgca tgacctcttg acgcgcggca tcgacgccaa cggcgaattg
     5641 cgcccacctc aggccgaggc accgcatctc tacgagtcgt caccgttggg ttggattccg
     5701 aatgagtggg gtcttgctcc tacagcaact cgctgccatc tgataaccaa aggcactacc
     5761 cctgcggcta atgagatgtg gcagggtggc gcgggaatta ggtttctgcg agtcgataat
     5821 ctttctttcg atggacaact ggatctagat gcaagcacgt ttcgagttag ccttgccacg
     5881 cacaaaggtt ttctggctcg ttcaagatgc cttgaaggtg atgtgctgac gaacatcgtt
     5941 ggcccacctc tagggaaact ggggcttgtt accaaagaaa ttggtgaggt caatattaat
     6001 caagcaattg cgttatttcg accaaccgaa caactactgc caaagttcct attaatctgg
     6061 cttagtagct caatctcgca gtcttggctg aggaaccgag ccaagcagac gtcgggacaa
     6121 gtgaatctga ccctcgctct atgccaggag cttcctctac ctcggatgac gatcaatgag
     6181 caacaggcaa tcgttgaccg agttgatgcc gcgcaggaac aaatctggtg tgaggaggaa
     6241 ctgatccgaa agatgcgact tgagaaatct ggccttatgg atgacctcct caccggccgc
     6301 gtccgcgtca agccgcagct ggcggaaacc aaacaagcag ggagcgcctg agtttaaact
     6361 gccttcgccg gtgaaattgc caagccttca gatcggtgac ctccggttca cgctccagcg
     6421 gagcgcgcgc cgcagaacta tgcagatcac cgtggagcgc agtggcgact tgatgctctg
     6481 cgcaccgccg gaggtggacg aggccgcgct gcgagcattc gtgctggaga agcgcttctg
     6541 gatctacacc aagctggccg agaaggaccg cttgcagcgc caggttccgc gcaaggaatt
     6601 cgtcggaggc gagggattct tgtatctcgg ccgcagccat cggctgaagg tggtcgatga
     6661 acagaatgtg ccactgaagt tgaatggagg ccgcttttgt ctgcgccgtg acgccctacc
     6721 cgccgcgcgc gagcatttca tccgctggta cggcgagcgt gccaaggcct ggctttcggg
     6781 gcgtgtagct gactaccagt cgcgaatgga ggtgacgcct gccggcgtca aggtgcagga
     6841 ccttggatat cgctggggtt cgtgtggcaa gggcgactgg ctgtacttcc actggaaggc
     6901 aatcctgctg ccggcgcgca tcgctgagta tgtcgtggtg catgagattg cccatctgca
     6961 tgagccgcac cacacgcctg cgttctggct tcgagtggag cgtgccatgc cggactatgc
     7021 gcaacgcaag gcctggctgg ccgagcatgg aatcgatgtt gaaggaatct aaagaacgat
     7081 ggctgactat ttcaccagtg actacttcaa gctgctgaac aagtggaagg ggcagaagcg
     7141 tgacgagtcc aaccccgagc agaaccgcgc ttatgaagat ctgaagaagg cctacgaggt
     7201 gacggaggcg tgggcggaca aggttaaggc cgagttgttc cctgtcgggc gcgtcgagat
     7261 tcgtaagcgc ccgaccaacc agggcaacaa ctttgccagc tacaactggg ccaaaatcta
     7321 cccttcatct gaggcgccga aagagttggc ttacacagtt ggcatcggcg ccgatgacgg
     7381 cttcgtagtc aagatcgata ccgttgaagc ttggcactgg ccgtcgtttt acaacgtcgt
     7441 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     7501 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     7561 aatggcgaat ggcgataagc tagcttcacg ctgccgcaag cactcagggc gcaagggctg
     7621 ctaaaggaag cggaacacgt agaaagccag tccgcagaaa cggtgctgac c
//
