LOCUS       sequence-420123-        5653 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pkk223 with Marinosulfonomonas MutY insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5653)
  AUTHORS   Utzman PH, Mays VP, Miller BC, Fairbanks MC, Brazelton WJ, Horvath 
            MP
  TITLE     Metagenome mining and functional analysis reveal oxidized guanine 
            DNA repair at the Lost City Hydrothermal Field.
  JOURNAL   PLoS One. 2024 May 8;19(5):e0284642. doi: 
            10.1371/journal.pone.0284642. eCollection 2024.
  PUBMED    38718041
REFERENCE   2  (bases 1 to 5653)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2024-05-8"; volume: "19(5):e0284642. doi"; pages: " 
            10.1371/journal.pone.0284642. eCollection 2024"
COMMENT     Sequence Label: pkk223_MsMutY
FEATURES             Location/Qualifiers
     source          1..5653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1144..1161)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1314..1400
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1492..1519
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1538..1629
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1630..1698,1699..2490)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1848..1867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2661..3249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3150..3169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3403..3420
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3435..3575
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3501..3520)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3661..3683
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3677..3868)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5326..5345
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(5592..5597,5598..5613,5614..5620)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5628..5644
                     /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)."
     primer_bind     join(5633..5653,1..2)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 ggaaacagaa ttcggtgacc atgcgcgact accgcgacaa ctccgaactg gggaccgtcc
       61 ttctggggtg gtacgatgct aacgcgagaa gcttaccctg gcgtgtagcc ccagcacgtt
      121 cgaaagcagg ggctttgccg gatccatacc gtatttggat gagcgaagtc atgctgcagc
      181 aaacgactgt cgctgcagtc gctaaatatt ttcacgcgtt tacgactcgc tggccaacca
      241 ttatcgcctt ggcatctgcc aatgacgatg atgttatggg acagtgggca ggcctgggtt
      301 actatgctcg tgcacgcaac cttttgaagt gtgcacgcgt ggtagtagcg gattatggcg
      361 gcaatttccc cgacacacgc gatgagttgc ttaaacttcc tggcattgga ccctatacgg
      421 cggcagctat tgcaagtatc gccttcggtc gttcagaaac agtgatggat gcgaatatcg
      481 aacgcgtgat ggcacgtatg catgacatcc attcgccgtt gcccagtgct aagcctgaac
      541 ttcttgcatg cgcggcagcc cttaccccgg atttccgtgc cggcgattat gctcaagctg
      601 taatggactt aggcgccacc atttgcacgc cgaagtctgc atcttgtgga gtgtgcccct
      661 ggtccttcgc atgcctagcc cgtatcaatg gtacagctgc cgtcttaccg aaaaagactc
      721 ctaaacccgt caagccaaca cgctacggaa tcgcgtacat cgcaagacgt ggggacggtg
      781 cgtggttgtt agaacgccgt cctgataaag gtttactggg aggaatgctg ggctggcccg
      841 gggctcagtg gggcgatcag gcggtagaca agccgcctgt ttcagctgat tggcaagacc
      901 ccggtgccga agtgcgccat tcatttacgc actttcatct gcgtttatcg ttacgcatta
      961 gcgtcgtcgg taatgcgaaa ccaaccgcag ggcacttcat cgcggatgca gattttgacc
     1021 ccgagacact acccaccgta atgcgtaaag catatcgtat tgcacgcaca aagtttgacg
     1081 gcgacgcgaa cggttgactg cagccaagct tggctgtttt ggcggatgag agaagatttt
     1141 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     1201 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     1261 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     1321 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     1381 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     1441 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     1501 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
     1561 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     1621 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     1681 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     1741 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     1801 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     1861 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     1921 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1981 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2041 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2101 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2161 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2221 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2281 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2341 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     2401 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     2461 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     2521 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     2581 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2641 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2701 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2761 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2821 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2881 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2941 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3001 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3061 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3121 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3181 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3241 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3301 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3361 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3421 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3481 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     3541 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     3601 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3661 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3721 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     3781 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     3841 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     3901 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     3961 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     4021 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     4081 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     4141 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     4201 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     4261 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     4321 tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg
     4381 agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc
     4441 gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc
     4501 ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac
     4561 aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc
     4621 ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag
     4681 cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa
     4741 cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc
     4801 tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     4861 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     4921 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     4981 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     5041 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     5101 tgggttgaag gctctcaagg gcatcggtcg acgctctccc ttatgcgact cctgcattag
     5161 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     5221 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     5281 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     5341 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     5401 gaggatccgg gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg
     5461 gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg
     5521 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
     5581 gaaatgagct gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata
     5641 acaatttcac aca
//
