LOCUS       sequence-135533-        5648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Acetobacter aceti 1023 Est05, C-terminal His6 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5648)
  AUTHORS   Kappock
  TITLE     Additional Acetobacter aceti 1023 hydrolase/transferase enzymes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5648)
  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            "Unpublished"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: pJK406
FEATURES             Location/Qualifiers
     source          1..5648
                     /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             2229..2251
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2282..2301)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2283..2301)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2810..3001
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2995..3017)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3103..3245
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3158..3177
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3260..3277)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3431..4019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3511..3530)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4190..4981,4982..5050))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4813..4832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5051..5155)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5182..5637)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5319..5340)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5531..5550
                     /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 gagtggatgt accggaggtg
      181 ttgtaggctg atattcaccg ccaatgcgtg tgatattctg tttgctcata tcacctgttt
      241 ctggaatccc tggggtaagg tgggcaaaat gctccaacag aatgcggcgt acttctgcaa
      301 tggtaatggg gttagattgg gtggaatgcc cggagtgccg cacaaccagt tcagaatcag
      361 catagggcac gtgggcgctt ttatatttta caaccccatc atctgccgta gctaaggggc
      421 catctcccaa cactggaata atggaatgtg tgcgtacctc tggcacaaca ggtacagaag
      481 ccagagagcg aataaaggcg ctatgggggg tcatgccata tacacttccc atacgccacg
      541 ggctcatatc tacccgtatg gagccagttt ctccggccac aacctgttgc ataacttcgg
      601 tcaggctcac aggaaatgtc accatacgtc caacaagatg cgctaaggaa agtgcagcca
      661 gataactgcc gtgttgtggt gtggaaataa aaacaacctg ttggacttct ggcaatggtt
      721 ttggaaatag actttcctgc aagaagctgc gggctttagg ggtaaggcgt aaactgttga
      781 atgggtgatg tgtcaggccg ttccaaagtt tatctttggc gtctatcact tgcatttttg
      841 ccaacagtcc cccttggcta tggccaataa gcaccatatg ccccaaagca ggatcggctt
      901 gtgtaccacc attgttttgg atggcggatt ccaatgcctc acggagttgc aaggcagagt
      961 agggaatggg gtttcctgta gcgtaggaaa agaaccagaa ttcgaaatgg gagcgaattt
     1021 ccggatcttc caacaggtca ttcaccatat ctgcccaacg gccagggctg gaagcagtgc
     1081 cgtggataag cacgacagga atgcacccaa agcggtgagg ttctatggca ataagttgcg
     1141 gtttgcggcc attgtcaaaa aaacgaccat ccagaaaacc cttgtattcg ctagaccagt
     1201 ctacagcatt gcttaggctc aatgcgcgcg aagctgtttg atcatactgc agggggataa
     1261 gtgtatggtt ggttgtgaga atatggccat tttcatccag agctgtcaga gtgaaatggc
     1321 ctgagatatg gttagaaaga acttgcctgc gtggatcagg gatttccata aatagactta
     1381 caggaatgcg cagtttatct gtgactgtaa aagaagaagg tgcacgccca tctacatgcg
     1441 gaatggcagc aaggggctct cctaatccca tatggctata aatgttttta acgccgttta
     1501 cggcaaggcg tgccgtggga cgaaaatctg tgagttcata gccatgccac atatgttctg
     1561 ctgggttggc tgaaaggtct aaatcgccaa atggaagtac ccaatgttga gcattgatat
     1621 tggcaggaga acccaaggct tcactcaggc tgagcatata aatatcacat gcttgccgaa
     1681 aatgcggatc atatggatct ggatgttctg tggttttatc ttctgggttg aggtaggcat
     1741 acgcgtataa ggcagcagcc atgaaatctg tgcgcgattt gtgttttttg ccccgcaggt
     1801 aacttagctc ggccaaagca aataactggt cgtttaaatc aggtgtgtaa aattggtctt
     1861 gcgtgctttt gcgcaaggta tctatggcag cgctggggtg tttttgccat tcacccagca
     1921 aattttggcg ttccagaaca atacgagtgg tgttgctaag tgcagttcca ttcagggccg
     1981 tatggttacg gtcctgataa gccttggtca ggctgagttg ttgaactgaa acaggaggcg
     2041 cacaacctgc aagagctaaa aagacagcaa cgcaacaaga tttatgattt aattttttaa
     2101 taaaactcat tgaaaaaaca aacattatgc tttccgcaaa acatatccaa acgttaaaat
     2161 gtggggtgga acttacgatg ttcattacaa tgtggcaaat ggaataaagg aatgtcatcc
     2221 atggtatatc tccttcttaa agttaaacaa aattatttct agagggaaac cgttgtggtc
     2281 tccctatagt gagtcgtatt aatttcgcgg gatcgagatc tcgggcagcg ttgggtcctg
     2341 gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg
     2401 ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg
     2461 caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag
     2521 tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg
     2581 ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg
     2641 agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc
     2701 cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat
     2761 cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca
     2821 ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga
     2881 gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca
     2941 cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     3001 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3061 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     3121 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
     3181 agagtgcacc atatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     3241 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3301 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3361 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3421 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3481 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3541 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3601 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3661 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3721 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3781 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3841 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3901 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3961 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4021 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4081 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4141 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4201 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4261 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4321 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4381 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4441 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4501 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4561 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4621 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4681 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4741 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4801 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4861 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4921 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4981 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5041 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5101 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5161 ttccccgaaa agtgccacct gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa
     5221 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5281 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5341 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5401 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5461 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5521 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5581 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     5641 attcgcca
//
