LOCUS       sequence-131414-        4694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Production of Acetobacter aceti 1023 alanine racemase (alr) with 
            C-terminal His tag (AaAlrH6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4694)
  AUTHORS   Francois JA, Kappock TJ
  TITLE     Alanine racemase from the acidophile Acetobacter aceti.
  JOURNAL   Protein Expr Purif. 2007 Jan;51(1):39-48. Epub 2006 Jun 6.
  PUBMED    16843006
REFERENCE   2  (bases 1 to 4694)
  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: "Protein 
            Expr Purif."; date: "2007-01"; volume: "51(1)"; pages: "39-48. Epub 
            2006 Jun 6"
COMMENT     Sequence Label: pJK301
FEATURES             Location/Qualifiers
     source          1..4694
                     /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             1275..1297
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1328..1347)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1329..1347)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1856..2047
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2041..2063)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2149..2291
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2204..2223
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2306..2323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2477..3065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2557..2576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3236..4027,4028..4096))
                     /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     3859..3878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4097..4201)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4228..4683)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4365..4386)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4577..4596
                     /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 gagggcagcc gttgcataaa
      181 cgcgatgata acgccgcccc agagatgtca gtacttcgta gcctattgtg ccttctgctt
      241 tggccacagc atctacgccg taagactcat tcaaaagttc caccatgcta tcttcttcca
      301 gcagcccttc gggcactgcg gaaagatcaa ctgtcatgga atccatagag attcggccca
      361 gaattggtag ccgatatcct ttataccaag cgcatcctgc agcagcgccc gcacgggcaa
      421 atccatcagc atatccaacc gcaatggtcg caacacggca tgggccagtc gcttgccacg
      481 taagcccata tcccacccga tcccctttgg ataaagtgcg aatttgcaga atatgcgttt
      541 gcaaccccac aacaggctta agtggattaa caccatcatt ctgcggcgct acgccataca
      601 aagcggcacc agggcgcacg agatcaaaat gaaatggttg ccccagaaaa attccagaag
      661 atgcagaaag acttaaagct gcctttggca acatgcgtgc atagttctca aaccgttcac
      721 gctgcatggc attggctgcg ttttctggcg tatctgcaca ggccagatga ctcataacca
      781 gactgacaga taacccctcc aaaagttcag gagtggccat cagtttatgg atatcagcct
      841 ctgaaaggcc aaaacgagac atacccgtat ccagttgaat ggccacatcc agcggcttat
      901 ccattttggc tgccagatcc cgcaaaacag ccacctgttt taggctattt aaaactggcc
      961 gcaaattatt gcgtacgcaa acctcagcag catccggccg tggaccatgc aacacagtta
     1021 ctgatgtgtg gggtgaaaga tatggcctaa gggctagccc ttcatccaca tgtgcgacaa
     1081 aaaaatgttt tgcgccaacc tgttccaaaa tgggcgccac ctgtgcaacc ccaagcccat
     1141 aagcatcggc cttaactact gcggcgcaaa ctgttttgcc cagtttgttc acaagatagc
     1201 ggtaattttc ggcaatagcg gaaagattta tcgtcaacac agcccctgcg cgcgatgctg
     1261 gccacatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag ggaaaccgtt
     1321 gtggtctccc tatagtgagt cgtattaatt tcgcgggatc gagatctcgg gcagcgttgg
     1381 gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg
     1441 gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg
     1501 ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc
     1561 gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc
     1621 aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg
     1681 accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca
     1741 acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg
     1801 tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac
     1861 caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct
     1921 tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca
     1981 cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa
     2041 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     2101 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     2161 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     2221 gtactgagag tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2281 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2341 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2401 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2461 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2521 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2581 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2641 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2701 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2761 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2821 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2881 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     2941 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3001 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3061 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3121 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3181 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3241 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3301 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3361 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3421 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3481 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3541 ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3601 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3661 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3721 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3781 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3841 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3901 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3961 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4021 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4081 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4141 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4201 gcacatttcc ccgaaaagtg ccacctgaaa ttgtaaacgt taatattttg ttaaaattcg
     4261 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4321 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4381 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4441 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4501 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4561 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4621 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4681 cgtcccattc gcca
//
