LOCUS       sequence-136191-        6078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Escherichia coli adenylate kinase (PlsA), N-terminal His6 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6078)
  AUTHORS   Schultheisz HL, Szymczyna BR, Scott LG, Williamson JR
  TITLE     Pathway engineered enzymatic de novo purine nucleotide synthesis.
  JOURNAL   ACS Chem Biol. 2008 Aug 15;3(8):499-511. doi: 10.1021/cb800066p.
  PUBMED    18707057
REFERENCE   2  (bases 1 to 6078)
  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"; doi: "10"; journalName: 
            "ACS Chem Biol"; date: "2008-08-15- 15"; volume: "3"; issue: "8"; 
            pages: "499-511"
COMMENT     Sequence Label: pET22-HT/plsA
FEATURES             Location/Qualifiers
     source          1..6078
                     /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"
     CDS             complement(830..850)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(854..871)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             882..904
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    919..943
                     /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(943..962)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(944..962)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1046..1065)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1271..1348
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1349..2431
                     /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(1368..1387)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3240..3431
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3425..3447)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3533..3675
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3588..3607
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3690..3707)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3861..4449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3941..3960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4620..5411,5412..5480))
                     /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     5243..5262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5481..5585)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5612..6067)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5749..5770)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5961..5980
                     /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 gcaagctttt
      181 agccgaggat tttttccaga tcagcgcgaa cttcagcaac cggcttggtg ccgtcaactt
      241 tcgcgtattt ggtgttaccc gcttccgctt ctttggagta gtagccgatc agcggtgcag
      301 tcatctgatg gtattcaacc agacgtttac gcacggtttc ttcctgatcg tctttacggg
      361 tagtcagttc ttcaccggta acgtcgtctt tgccttctac tttcggcgga ttgaatttaa
      421 cgtgataaac acgaccagac ggcgcatgaa cgcggcggcc tacgatacga tcaacaatca
      481 gttcgtccgg tacgtcgaat tccagaacgt aatcaacatt gatgcccgct tctttcatcg
      541 cgtctgcctg cggaatggta cgcgggaagc cgtccaacag gaaaccatta cggcagtctt
      601 cctgagcaat gcgctcttta accagcgcga tcaccagttc gtcggtgacc agtttgccag
      661 catccataat gtcttttgct tgtttaccca gctcggagcc agatttgacc gcagcacgca
      721 gcatatcgcc agtggagatt tgcggaatac catatttctc catgatgaac tgagcctgag
      781 tccctttccc cgcgcccgga gcgccaagca gaatgatacg catggatccg ccctgaaaat
      841 acaggttttc gccgtgatga tgatgatgat gcatatgtat atctccttct taaagttaaa
      901 caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat
      961 taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc
     1021 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
     1081 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
     1141 gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc
     1201 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
     1261 cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
     1321 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     1381 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     1441 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     1501 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     1561 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     1621 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     1681 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     1741 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     1801 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     1861 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     1921 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     1981 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     2041 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     2101 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     2161 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     2221 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     2281 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     2341 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     2401 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
     2461 ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     2521 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     2581 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
     2641 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
     2701 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
     2761 gcatggcggc cccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct
     2821 ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg
     2881 actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg
     2941 tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca
     3001 tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc
     3061 attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct
     3121 cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct
     3181 ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag
     3241 tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa
     3301 cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc
     3361 ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg
     3421 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     3481 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     3541 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     3601 gattgtactg agagtgcacc atatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     3661 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3721 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3781 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3841 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3901 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3961 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4021 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4081 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4141 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4201 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4261 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     4321 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4381 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4441 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4501 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4561 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4621 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4681 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4741 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4801 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4861 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4921 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4981 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5041 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5101 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5161 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5221 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5281 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5341 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5401 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5461 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5521 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5581 ccgcgcacat ttccccgaaa agtgccacct gaaattgtaa acgttaatat tttgttaaaa
     5641 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5701 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5761 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5821 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5881 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5941 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6001 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6061 ggcgcgtccc attcgcca
//
