LOCUS       sequence-139780-        9306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces formylglycinamide ribonucleotide amidotransferase, 
            N-terminal His6 tag (H6EcPurL).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9306)
  AUTHORS   Kappock
  TITLE     Purine biosynthesis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9306)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pJK556
FEATURES             Location/Qualifiers
     source          1..9306
                     /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(4058..4078)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(4082..4099)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4110..4132
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4147..4171
                     /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(4171..4190)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4172..4190)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4274..4293)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4499..4576
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4577..5659
                     /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(4596..4615)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             6468..6659
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6653..6675)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6761..6903
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6816..6835
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6918..6935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7089..7677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7169..7188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7848..8639,8640..8708))
                     /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     8471..8490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8709..8813)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8840..9295)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8977..8998)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9189..9208
                     /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 gagttacccc aactgcttac
      181 gcgcattgcg gaaaatgcgc atccatgggc catcctcgcc ccagttttcc ggatgccagg
      241 agttgctgac agtacggaaa acacgttccg ggtgcggcat cataatggtg actcgaccac
      301 tttcagtcgt aaccgacgta ataccgttcg gtgaaccgtt cgggttagcc gggtaggttt
      361 cagtgacttt gccgaagtta tcgacatagc gcagtgccac cagccctttg ctttccagtg
      421 ccgccagatg cgccgcatca cgcacttcca cgcgcccttc accatgagag acagcaatcg
      481 gcatttgcga gcccaccatc ccctgcaaca gcagagacgg gctttgggtt acttcaacca
      541 ggctgaaacg cgcttcaaag cgatcggagg tattgcgcac aaaacgtggc cacaactcac
      601 tacctgggat cagttcgcgc aggttagaca tcatctggca accgttacat acccccagcg
      661 ccagcgtttg cggacggtgg aagaaggttg caaactcatc gcgtacacgg tcattgaaca
      721 ggattgactt cgcccaacct tcaccggcac ccagcacatc accgtaggag aaaccaccgc
      781 acgcgaccag ggcgtggaaa tcttccaggc ccgtgcgtcc ggtcagcagg tcactcatat
      841 gcacgtcgat agcatcaaag cctgcacggt ggaaagctgc cgccatttca acatgcgagt
      901 tcacgccctg ctcacgcagt acggcaactt tcggacgtgc gccagtggca atatacggtg
      961 ctgccacatc ttcgttgata tcgaacgaca gttttacatt caggcccgga tcggcgtcgt
     1021 tagatttcgc ctgatgctcc tgatcggcac actccgggtt gtcacgcagg cgctgcatct
     1081 gccaggtagt ttctgcccac cagacacgca acgtggtgcg gctttcgctg aatacagtct
     1141 gcccgttggc ggtaatcaca aaacggtcac cggaaaccgc ctgccctaca taatggacac
     1201 aatcagcaag cccatgctgt gccagtacgg actcgaccgc ttcacggtca gcggcacgaa
     1261 cctgaatcac cgcacccagt tcttcgttaa acaacgccgc caggcgatcg tcgcccagag
     1321 aggcaatatc cgcgttaatg ccgcaatgac cagcaaaggc catttccgcc agcgttacca
     1381 gcaggccgcc atcagagcgg tcgtggtatg cgagcagctt acgctgtgca accagcgcct
     1441 gaatcgcgtc atagaagcct ttcagttgcg cgacgtcgcg tacatctgcc ggtttgtcgc
     1501 caagctgacg ataaacctgc gccagcgccg ttgcgcccag cgcgttattg cctttgccca
     1561 aatcaatcag cagcagtgcg ttatcttcgg tagaaagctg tggcgtgatg gtgtgacgca
     1621 catcttccac gcgggcaaat gcagaaatca ccagcgacag cggcgacgtc atttcgcgct
     1681 cttcgttacc ttcctgccag cgggttttca tcgacataga gtctttaccc accgggatcg
     1741 tcaggcccag cgccggacaa agctcttcgc ccacggcttt aacggcttca tacaggcccg
     1801 catcttcacc agggtggcct gccgccgcca tccagttggc ggaaagttta atgcgtttga
     1861 tatcgccaat ctgtgttgcg gcaatattgg ttaacgcttc accgaccgcc agacgggcag
     1921 aggcggcgaa atccagcagc gcaaccggcg cacgctcgcc aatcgccatc gcttcaccgt
     1981 agtagctgtc gaggctggcg gtagtgaccg cgcagttagc gaccggcacc tgccacggac
     2041 ccaccatctg atcgcgcgct accatgccag ttacgctgcg gtcaccgatg gtcaccagga
     2101 aagttttctc cgccacggtc ggcagatgca gtacgcgttt caccgcgtct gcaatggtga
     2161 tcccttcacg gaccagcgca tcgcccttcg ctttcagcgt ttgtacatcg cgggtcatct
     2221 tcggcgtttt accaagcagg acgtccagcg gcagatcgat cggctgatta tcaaaatgac
     2281 gatcgtgcag agaaagatgc agttcttcgg tcgcttcacc aatcaccgcg tagggtgcgc
     2341 gctcacgctt acacagttcg tcaaacagcg gtaattgatc ggcagcaacc gccagcacgt
     2401 agcgttcctg ggattcgtta caccagattt ccagcgggct catgcctggc tcgtcgctca
     2461 gaatgtcgcg cagttcaaat ttaccgccac gcccgccgtc gctcaccagt tccggcatag
     2521 cgttagaaag accgcctgcg ccaacgtcgt ggataaacag tattgggttg gcatcaccaa
     2581 gctgccagca acggtcaatc acttcctggc agcgacgctc catctccggg ttgtcgcgct
     2641 gtacggaagc aaagtcgagg tcggcatcag actgaccaga cgccatggaa gacgccgcgc
     2701 cgccgccaag accgatgttc attgccgggc cgccgagaac gaccagcttc gcgccaacgt
     2761 tgatctcgcc tttttgtacg tgatcggcgc gaatgttgcc gatcccgccc gccagcatga
     2821 tcggtttgtg ataaccgcgc agctcttcgc cgttgtggct gttcactttt tcttcataag
     2881 tacggaagta gccgttcagt gccggacgac caaattcgtt gttaaacgcc gcaccgccca
     2941 gcgggccttc ggtcatgatg tccagcgcgg tgacaatgcg ctcaggctta ccgaaatctt
     3001 cttcccacgg ctgttcgaag ccaggaattc gcaggttgga aacggagaaa ccaaccaaac
     3061 ccgctttcgg cttcgcgccg cgcccggtgg caccttcatc gcggatttca ccgccggaac
     3121 cggtcgccgc gcccggccac ggagaaatcg ccgtcgggtg gttgtgagtt tcgactttca
     3181 tcagaatatg cgccggttcc tgatggaaat catagcggcc cgtttcgtgg tcagcaaagt
     3241 agcggcccac ttcagaacct tccattacgg cggcgttatc tttataagca gagagaacgt
     3301 gatctggcgt ggtttcgaaa gtatttttga tcatcctgaa cagcgatttc ggctgttgtt
     3361 caccgtcgat aatccagtcg gcgttaaaaa ttttgtggcg gcagtgctca gagttcgcct
     3421 gagcgaacat atacagctcg atgtcgttcg ggttacgacc aagctttgtg aaagcatcct
     3481 gcagatagtc aatttcatct tccgccagag ccaggccaag acgcaggtta gcgtcgatca
     3541 gcgcctgacg gccctgcccc agcaaatcaa cgcttgtgac cggagtcggt tgatggtggg
     3601 caaacaactg ctctgcatca tctaaatcaa aaaagaccgt ttccatcatg cggtcgtgca
     3661 gttcagcggt aacctgctgc cattgttcat tggtcagcgt accggcttct atatagtaag
     3721 caacgccgcg ctcaaggcgg tttacctgtt gtagcccgca gttatgggca atatcggtcg
     3781 ctttcgaaga ccagggagag atggtgccag gacgcggggt caccagcagg agtttgcctt
     3841 gcggggcgtg gctggcgagt gccggaccat atttcagcag gcgttcaagt tgtgcgtgct
     3901 catcatcgtt taacggcgca ttgaggtcag caaaatggac atactcggcg taaatattgt
     3961 gaaccgggag cctggcagcc tgaaaacgtg ccagcagttt gttgattcgg aatgccgaca
     4021 gtgcaggcga accacgcaga atttccatca tagatccgcc ctgaaaatac aggttttcgc
     4081 cgtgatgatg atgatgatgc atatgtatat ctccttctta aagttaaaca aaattatttc
     4141 tagaggggaa ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg
     4201 atcgagatct cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     4261 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
     4321 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
     4381 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
     4441 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga
     4501 caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     4561 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     4621 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     4681 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     4741 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     4801 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     4861 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     4921 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     4981 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     5041 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     5101 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     5161 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     5221 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     5281 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     5341 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
     5401 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     5461 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     5521 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
     5581 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     5641 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     5701 cgggatctcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     5761 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     5821 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     5881 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     5941 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccc
     6001 cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc
     6061 ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca
     6121 aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc
     6181 tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct
     6241 gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag
     6301 tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca
     6361 gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg
     6421 gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg
     6481 aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga
     6541 aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg
     6601 ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     6661 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     6721 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     6781 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     6841 agtgcaccat atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     6901 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6961 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7021 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7081 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7141 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7201 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7261 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7321 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7381 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7441 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7501 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     7561 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7621 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7681 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7741 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7801 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7861 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7921 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7981 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     8041 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8101 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8161 gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8221 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8281 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8341 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8401 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8461 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8521 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8581 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8641 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8701 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8761 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8821 ccccgaaaag tgccacctga aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     8881 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     8941 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     9001 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     9061 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     9121 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     9181 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     9241 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     9301 tcgcca
//
