LOCUS       sequence-136192-        6400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Escherichia coli phosphoribosylpyrophosphate synthetase 
            (PrsA), N-terminal His6 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6400)
  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 6400)
  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/prsA
FEATURES             Location/Qualifiers
     source          1..6400
                     /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(1152..1172)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1176..1193)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1204..1226
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1241..1265
                     /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(1265..1284)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1266..1284)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1368..1387)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1593..1670
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1671..2753
                     /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(1690..1709)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3562..3753
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3747..3769)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3855..3997
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3910..3929
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4012..4029)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4183..4771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4263..4282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4942..5733,5734..5802))
                     /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     5565..5584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5803..5907)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5934..6389)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6071..6092)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6283..6302
                     /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 cgaattctta gtgttcgaac atggcagaga tcgattcttc gttgctgata
      241 cgacgaatcg cttcggccag catacctgac agggtcagag tacgcacgtt cggcagtgat
      301 ttgatttcat cgctcagcgg aatggtatcg cagacaacga cttcatcaat tacagagtta
      361 cgcaggttgt tcgccgcgtt gccagagaag atcgggtgag tcgcgtacgc aaatacacgt
      421 ttagcaccac gttctttcag agcttcagca gctttacaca gcgtaccgcc agtgtcgatc
      481 atatcatcga ccagtacgca gtcacgacct gcaacgtcac cgatgatatg catcacctgg
      541 gaaacgttcg cacgcggacg acgtttgtcg atgattgcca tatcggtatc gttcagcagc
      601 ttagcgatag cgcgggcacg cacaacgccg ccgatgtccg gagaaaccac aattgggtta
      661 tccagattca gctgcagcat gtcttccagc aggatcgggc taccaaatac gttatcaacc
      721 ggaacgtcga agaaaccctg aatctgttca gcgtgcagat ccactgtcag cacacggtca
      781 acaccgacgc tggagaggaa gtctgcaacc actttcgcag tgattggtac acgagcggaa
      841 cggacgcgac ggtcctggcg cgcatagcca aagtagggga taacagcggt gatacgacct
      901 gcggaagcac gacgcagggc atcaaccata acgactaatt ccatcaggtt gtcgttagta
      961 ggggcacaag tggactggat gatgaaaata tcaccaccgc gtacattttc attaatttgt
     1021 acgctgactt cgccatcgct aaagcgacct acagcggcgt cgccgagtga agtgtacagg
     1081 cggttggcaa tacgttgtgc tagttccggg gtggcgttac cagcaaaaag cttcatatca
     1141 ggcacggatc cgccctgaaa atacaggttt tcgccgtgat gatgatgatg atgcatatgt
     1201 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
     1261 attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg
     1321 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
     1381 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
     1441 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
     1501 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
     1561 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc
     1621 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     1681 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     1741 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     1801 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     1861 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     1921 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     1981 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     2041 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     2101 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     2161 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     2221 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     2281 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     2341 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     2401 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     2461 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     2521 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     2581 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     2641 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     2701 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     2761 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
     2821 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     2881 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     2941 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     3001 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     3061 gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg
     3121 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     3181 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     3241 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     3301 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     3361 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     3421 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     3481 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     3541 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     3601 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     3661 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
     3721 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3781 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3841 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     3901 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat
     3961 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     4021 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4081 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4141 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4201 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4261 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4321 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4381 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4441 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4501 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4561 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4621 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4681 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4741 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4801 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4861 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4921 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4981 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5041 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5101 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5161 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5221 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
     5281 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5341 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5401 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5461 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5521 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5581 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5641 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5701 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5761 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5821 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5881 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt
     5941 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     6001 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     6061 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6121 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6181 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6241 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6301 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6361 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
