LOCUS       sequence-133865-        7298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Acetobacter aceti 1023 small hypothetical protein with 
            N-terminal His6 tag (H6AaOrfX); N5-carboxyaminoimidazole 
            ribonucleotide mutase (AaPurE1); NCAIR synthetase (AaPurK).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7298)
  AUTHORS   Kappock
  TITLE     Additional purEK associated constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7298)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pJK540
FEATURES             Location/Qualifiers
     source          1..7298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1362..1370
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(2172..2189)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2199..2216)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2235..2257
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2272..2296
                     /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     complement(2296..2315)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2297..2315)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2399..2418)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2624..2701
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2702..3784
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2721..2740)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4593..4784
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4778..4800)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4886..5028
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4941..4960
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5043..5060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5214..5802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5294..5313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5913..5932)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5924..6739
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             5924..6739
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5997..6016)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6832..7287)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6969..6990)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7181..7200
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattctta actggcagat tcttcatctt ccacagcggc cgttttggtg
      241 gccaatgggc ccaaggcatc ctgaataccc aaattgcctg gcagcgcacc gtaggggaac
      301 agaatattta catgccccat tttacggccc actctggctt ctgccttgcc atatagatgc
      361 ggaataaagc cctgcgtggc cataatctgc ggcagcagcg ccatatcttc cggcccgacc
      421 agatttttca tgattgcatc agaatggcgc acagcgtgcg gcaaaggcag ccccgcaact
      481 gcacggatat gcatgtcaaa ttggtcctgc gggcaggcat tcatggtcca atggccagaa
      541 ttatgggggc gcggggccat ttcattcacc agcaggttgc cggtgcaatc aacaaacatt
      601 tcaacgccca gaataccaac cagctccagc ttttctgcaa tggtggtggc tagttttatg
      661 gcctgcgtgg caatatctgg gtcaatgggg gcgggggcta atgttagatc cagaataccg
      721 ttccagtgcc ggttcagcgc cgggtcaaag caccttaccg tgccatctgc accacgcgcc
      781 accataacac taatctcgca agcaaaatcc accatacctt ctgcaaccag aggatgtggt
      841 gccagagcgt tgaaagcctc ttccagatca tggatgtgat gcacacggcg ctggcctttg
      901 ccatcatatc cattgcgagt ggttttgaga atcagcggca aacccacttt ttctgcggct
      961 tcgtgtagcg ttttgacatc ggtaacagga tgccatggcg cgagtggaat acccgcagaa
     1021 gcaaaaaaag ttttttctgc caatcggtcc tgactgatgc gcagaatatg ccccgaagga
     1081 tgaacagggc agtgtttggc taggcggtcc agtgcatctg cgctgatgtt ttcaaattca
     1141 aaggtaacaa catcaacagc gcgggcaaag gcatccagtg ctgtcggatc atcatacttg
     1201 ccgcatgtta cggcgtgaga gacctgagcc gccgggccgt tggcctctgg cgtaagaata
     1261 tgcgtgcgaa aaccaaggcg cgcggcagca acagcagaca tgcggccaag ctggccaccc
     1321 cccacaatgc caataacggc attgggctta agcggagaat gtaaggaggt catttatctt
     1381 cggtaatggg agagttgggg acagaggccg tctgcaatgc gcgccatgtt tcaaggcgtg
     1441 cggcaagggc cgggttatac agtgcaagaa tggaggcggc cagcaaggcg gcatttttgg
     1501 cgccggatgc cccaatagcc aaagtgccca caggcacgcc gccgggcatc tgcacaatgg
     1561 aaagcaggct gtccatgcct ttaagagcgc gagattctac tggaacaccc agcacgggca
     1621 ggcgtgtcca tgccgcacac atacccggca gatgtgcagc tccaccggcc ccagcaataa
     1681 tcacattcag gccgcgttcg gctgccgtac gcgcataatc cgccaaccgg tctggcgtac
     1741 gatgtgctga aacaatgagg gtttcatgcg gaatttccag ctctgtcagc aatgcgtctg
     1801 catggcgcat ggtttcccag tctgactggc tgcccatgat aatgcccaca acgggggcag
     1861 aggctgcttt atcttctaaa gcactggaag cagagggcag gggggcggtt tcgctcacgg
     1921 tgggtatcta tccggttttt taaagtctgt aacggagagg ttcaggcaat gtcatcaggg
     1981 atcagaaggc tttcaacctt ggtaatctga tcttttagca gaagtttacg ttttttcagc
     2041 cgctggagat agagttggtc aataaaagta gtttcatgcg ccatgcggtt gataaccgta
     2101 tccagatctc tgtgttcact ccggagttca tgcagttggg caagcagtaa atcgcgatct
     2161 cgcagcatat ggctgccgcg cggcaccagg ccgctgctgt gatgatgatg atgatggctg
     2221 ctgcccatgg tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt
     2281 atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc
     2341 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
     2401 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     2461 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
     2521 ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc
     2581 ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg
     2641 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
     2701 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
     2761 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
     2821 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
     2881 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
     2941 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     3001 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
     3061 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     3121 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
     3181 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     3241 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
     3301 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
     3361 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
     3421 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
     3481 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc
     3541 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     3601 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     3661 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
     3721 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3781 gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga
     3841 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     3901 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     3961 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
     4021 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
     4081 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga
     4141 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
     4201 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
     4261 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
     4321 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
     4381 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt
     4441 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt
     4501 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca
     4561 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta
     4621 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
     4681 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
     4741 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4801 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4861 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     4921 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg
     4981 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     5041 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5101 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5161 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5221 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5281 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5341 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5401 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5461 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5521 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5581 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5641 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5701 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5761 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5821 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaac
     5881 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     5941 ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     6001 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
     6061 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     6121 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     6181 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc
     6241 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     6301 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     6361 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     6421 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     6481 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     6541 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     6601 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     6661 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     6721 gctcgatgag tttttctaag aattaattca tgagcggata catatttgaa tgtatttaga
     6781 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gaaattgtaa
     6841 acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     6901 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     6961 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     7021 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     7081 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     7141 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     7201 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     7261 cgcttaatgc gccgctacag ggcgcgtccc attcgcca
//
