LOCUS       sequence-131052-        5348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Acetobacter aceti 1023 PurE and PurK in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5348)
  AUTHORS   Constantine CZ, Starks CM, Mill CP, Ransome AE, Karpowicz SJ, 
            Francois JA, Goodman RA, Kappock TJ
  TITLE     Biochemical and structural studies of N5-carboxyaminoimidazole 
            ribonucleotide mutase from the acidophilic bacterium Acetobacter 
            aceti.
  JOURNAL   Biochemistry. 2006 Jul 11;45(27):8193-208.
  PUBMED    16819818
REFERENCE   2  (bases 1 to 5348)
  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: 
            "Biochemistry."; date: "2006-07-11"; volume: "45(27)"; pages: 
            "8193-208"
COMMENT     Sequence Label: pJK174
FEATURES             Location/Qualifiers
     source          1..5348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        494..598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(599..667,668..1459)
                     /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     complement(817..836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1630..2218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2119..2138
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2372..2389
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2404..2546
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2472..2491)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2632..2654
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2648..2839)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3348..3367
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3348..3366
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3398..3420
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             3979..3987
                     /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             5192..5209
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5262..5280)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5276..5323
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     3301 cccgtggcca ggacccaacg ctgcccgaga tctcgatccc gcgaaattaa tacgactcac
     3361 tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga
     3421 tatacatatg atgagcgaaa ccgcccccct gccctctgct tccagtgctt tagaagataa
     3481 agcagcctct gcccccgttg tgggcattat catgggcagc cagtcagact gggaaaccat
     3541 gcgccatgca gacgcattgc tgacagagct ggaaattccg catgaaaccc tcattgtttc
     3601 agcacatcgt acgccagacc ggttggcgga ttatgcgcgt acggcagccg aacgcggcct
     3661 gaatgtgatt attgctgggg ccggtggagc tgcacatctg ccgggtatgt gtgcggcatg
     3721 gacacgcctg cccgtgctgg gtgttccagt agaatctcgc gctcttaaag gcatggacag
     3781 cctgctttcc attgtgcaga tgcccggcgg cgtgcctgtg ggcactttgg ctattggggc
     3841 atccggcgcc aaaaatgccg ccttgctggc cgcctccatt cttgcactgt ataacccggc
     3901 ccttgccgca cgccttgaaa catggcgcgc attgcagacg gcctctgtcc ccaactctcc
     3961 cattaccgaa gataaatgac ctccttacat tctccgctta agcccaatgc cgttattggc
     4021 attgtggggg gtggccagct tggccgcatg tctgctgttg ctgccgcgcg ccttggtttt
     4081 cgcacgcata ttcttacgcc agaggccaac ggcccggcgg ctcaggtctc tcacgccgta
     4141 acatgcggca agtatgatga tccgacagca ctggatgcct ttgcccgcgc tgttgatgtt
     4201 gttacctttg aatttgaaaa catcagcgca gatgcactgg accgcctagc caaacactgc
     4261 cctgttcatc cttcggggca tattctgcgc atcagtcagg accgattggc agaaaaaact
     4321 ttttttgctt ctgcgggtat tccactcgcg ccatggcatc ctgttaccga tgtcaaaacg
     4381 ctacacgaag ccgcagaaaa agtgggtttg ccgctgattc tcaaaaccac tcgcaatgga
     4441 tatgatggca aaggccagcg ccgtgtgcat cacatccatg atctggaaga ggctttcaac
     4501 gctctggcac cacatcctct ggttgcagaa ggtatggtgg attttgcttg cgagattagt
     4561 gttatggtgg cgcgtggtgc agatggcacg gtaaggtgct ttgacccggc gctgaaccga
     4621 cactggaacg gtattctgga tctaacatta gcccccgccc ccattgaccc agatattgcc
     4681 acgcaggcca taaaactagc caccaccatt gcagaaaagc tggagctggt tggtattctg
     4741 ggcgttgaaa tgtttgttga ttgcaccggc aacctgctgg tgaatgaaat ggccccgcgc
     4801 ccccataatt ctggccattg gaccatgaat gcctgcccgc aggaccaatt tgacatgcat
     4861 atccgtgcag ttgcggggct gcctttgccg cacgctgtgc gccattctga tgcaatcatg
     4921 aaaaatctgg tcgggccgga agatatggcg ctgctgccgc agattatggc cacgcagggc
     4981 tttattccgc atctatatgg caaggcagaa gccagagtgg gccgtaaaat ggggcatgta
     5041 aatattctgt tcccctacgg tgcgctgcca ggcaatttgg gtattcagga tgccttgggc
     5101 ccattggcca ccaaaacggc cgctgtggaa gatgaagaat ctgccagtta agaattcgag
     5161 ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac caccaccact gagatccggc
     5221 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     5281 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     5341 atccggat
//
