LOCUS       sequence-138495-        6082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Escherichia coli guanylate kinase (SpoR), N-terminal His6 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6082)
  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 6082)
  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/spoR
FEATURES             Location/Qualifiers
     source          1..6082
                     /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(834..854)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(858..875)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             886..908
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    923..947
                     /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(947..966)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(948..966)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1050..1069)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1275..1352
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1353..2435
                     /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(1372..1391)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3244..3435
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3429..3451)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3537..3679
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3592..3611
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3694..3711)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3865..4453)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3945..3964)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4624..5415,5416..5484))
                     /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     5247..5266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5485..5589)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5616..6071)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5753..5774)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5965..5984
                     /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 cgaattctca gtctgccaac aatttgctga ttaaagcgtc atgacgctgc
      241 ttttggcggc tcatgcgcag acgttcggcg cgaataatgg tcttcaaatc ggtcaacgcg
      301 gtatcgaagt catcattcac aatcagataa tcatattcgg cgtaatggct catttctgca
      361 acagcttgcg ccatacgctt tgcaatgacc tcttcgctgt cctgaccgcg accgcgtaga
      421 cggcggtcca gttcaatttt ggacggcggt aaaataaaga tactccgcgc gtgcggcatc
      481 ttctggcgaa tttgctgcgc gccctgccag tcgatatcga gaaaaacatc gacaccggtc
      541 gccagtactt gctcaatggc ctcacgcgaa gtgccatagt aattaccaaa aacttctgcg
      601 tgttcgagga acgcatctct gctaatcatt tctttaaatt catcatgatt aacaaagaaa
      661 taatgttcac cgtggacttc accaggacgc ggttggcgtg tggtgtgtga aacagaaact
      721 tgggtgtcat acaacggttg ggtttttaat aaagcctgaa tcaggctgga tttacccgcg
      781 ccactggggg cagaaacaat ataaagcgtg ccttgagcca tggatccgga tccgccctga
      841 aaatacaggt tttcgccgtg atgatgatga tgatgcatat gtatatctcc ttcttaaagt
      901 taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct atagtgagtc
      961 gtattaattt cgcgggatcg agatctcgat cctctacgcc ggacgcatcg tggccggcat
     1021 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
     1081 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
     1141 cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt
     1201 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
     1261 gcgtcgagat cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc
     1321 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
     1381 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     1441 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     1501 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     1561 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     1621 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     1681 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     1741 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     1801 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     1861 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     1921 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     1981 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
     2041 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
     2101 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
     2161 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     2221 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     2281 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
     2341 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     2401 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
     2461 ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc
     2521 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     2581 atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt
     2641 cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc
     2701 gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc
     2761 gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta
     2821 ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga
     2881 agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc
     2941 cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc
     3001 tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc
     3061 tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta
     3121 ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc
     3181 ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca
     3241 tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca
     3301 ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa
     3361 tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac
     3421 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     3481 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     3541 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag
     3601 agcagattgt actgagagtg caccatatat gcggtgtgaa ataccgcaca gatgcgtaag
     3661 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3721 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3781 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3841 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3901 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3961 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4021 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4081 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4141 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4201 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4261 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     4321 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4381 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4441 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4501 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4561 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4621 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4681 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4741 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4801 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4861 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4921 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4981 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5041 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5101 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5161 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5221 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5281 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5341 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5401 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5461 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5521 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5581 ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt gtaaacgtta atattttgtt
     5641 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5701 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5761 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5821 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5881 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5941 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     6001 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     6061 acagggcgcg tcccattcgc ca
//
