LOCUS       sequence-136194-        6132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Escherichia coli 5-aminoimidazole-4-N-succinocarboxamide 
            ribonucleotide synthetase (PurC), N-terminal His6 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6132)
  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 6132)
  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/purC
FEATURES             Location/Qualifiers
     source          1..6132
                     /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(884..904)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(908..925)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             936..958
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    973..997
                     /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(997..1016)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(998..1016)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1100..1119)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1325..1402
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1403..2485
                     /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(1422..1441)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3294..3485
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3479..3501)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3587..3729
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3642..3661
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3744..3761)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3915..4503)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3995..4014)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4674..5465,5466..5534))
                     /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     5297..5316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5535..5639)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5666..6121)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5803..5824)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6015..6034
                     /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 gagtcagtcc agctgtacac
      181 ccaggcggcg ggcgacggct tcataggcct cgatcaggcc accgaggctc tggcggaaac
      241 ggtctttgtc cattttctcc agtgtttctt tgtcccacag gcggctgccg tccggggaga
      301 actcatcgcc cagtaccact tcacctttgt acagaccaaa ctccagcttg aagtcgacca
      361 gaatcagacc agcatcatcg aacagttttt tcagcacgtc gttcgctttg taggtcagct
      421 ctttcatacg cgccaggttc tctttgctca cccagccaaa ggtttcgcag taagattcgt
      481 tgaccatcgg atcgtgcatg gcgtcgtttt tcaggaacag atcgaacagc ggcgggttca
      541 gctcaatacc ttcttcgatt ccaagacgtt tcaccagaga gccagcagca cggttacgca
      601 cgacacactc aaccggcacc atatccagct ttttcaccag acattcggta tcggagagca
      661 gacgctccat ttgagtcggg atacccgctt cagccagttt gctcataatg aagtagttga
      721 acttgttgtt caccatacct ttgcgatcaa actgctcaat gcgcgcgcca tcccctgctg
      781 acgtatcatt gcggaattcg agcaccaaca ggtccgggtt ttccgtgctg tatacggttt
      841 tcgctttacc acgatacaac tcagcttgct tttgcatgga tccgccctga aaatacaggt
      901 tttcgccgtg atgatgatga tgatgcatat gtatatctcc ttcttaaagt taaacaaaat
      961 tatttctaga ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt
     1021 cgcgggatcg agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
     1081 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
     1141 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
     1201 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
     1261 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat
     1321 cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     1381 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     1441 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     1501 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     1561 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     1621 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     1681 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     1741 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     1801 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     1861 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     1921 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     1981 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     2041 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     2101 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     2161 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     2221 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
     2281 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     2341 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     2401 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2461 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt
     2521 aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     2581 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     2641 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     2701 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     2761 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     2821 cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg
     2881 gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct
     2941 gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt
     3001 aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag
     3061 gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac
     3121 cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac
     3181 gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt
     3241 tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca
     3301 aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc
     3361 tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg
     3421 accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     3481 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     3541 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     3601 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     3661 actgagagtg caccatatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     3721 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3781 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3841 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3901 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3961 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4021 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4081 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4141 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4201 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4261 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4321 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     4381 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4441 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4501 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4561 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4621 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4681 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4741 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4801 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4861 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4921 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4981 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5041 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5101 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5161 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5221 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5281 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5341 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5401 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5461 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5521 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5581 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5641 acatttcccc gaaaagtgcc acctgaaatt gtaaacgtta atattttgtt aaaattcgcg
     5701 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5761 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5821 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5881 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5941 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     6001 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     6061 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     6121 tcccattcgc ca
//
