LOCUS       sequence-139433-        6070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Acetobacter aceti 1023 formylglycinamidine ribonucleotide 
            synthetase glutaminase subunit (AaPurQ).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6070)
  AUTHORS   Kappock
  TITLE     A multisubunit Gram-negative FGAM synthetase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6070)
  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            "Unpublished"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: pJK545
FEATURES             Location/Qualifiers
     source          1..6070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     RBS             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        147..165
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    166..190
                     /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)."
     CDS             1016..1060
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1098..1116)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1112..1159
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1196..1651
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1283..1302)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1493..1514
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1766..2557,2558..2626))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2389..2408
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2627..2719)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2800..3388
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3289..3308
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3542..3559
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3574..3716
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3642..3661)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3802..3824
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3818..4009)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4581..5663)
                     /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     5625..5644
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5664..5741)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5947..5966
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(6054..6070,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atggcaagct tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt
      121 attgtacacg gccgcataat cgaaattaat acgactcact ataggggaat tgtgagcgga
      181 taacaattcc ccatcttagt atattagtta agtataagaa ggagatatac atatgaaagc
      241 agcaattatc gtattccccg gcacaaaccg ggagcgcgat atggccattg ccctcaagcg
      301 cgttacaggg caggccccgc gcatgatctg gcaccatgaa acagatctgg gagataccga
      361 tctagtcgtg ctgccaggtg gtttcagcta tggcgattat ctgcgctgtg gtgccatggc
      421 agcgcattct cccatcatgg cggctatcaa agcctttgcc caaaagggtg gccatatttt
      481 gggtgtgtgc aacggcttcc agattttaac ggaaacaggg cttctgcccg gtgctctgtt
      541 gcgcaacgcc aacctgcgct ttctctctca ggattgctac ctgaaggtgg aagcagagaa
      601 cacacctttc actcacaagt ggaagaaggg tgatgttttc cgcacccctc tggcccatgg
      661 tgatggcaat tacacggcag atatcgccac gctggaacgg cttgaaggtg aaggtcttgt
      721 ggctttccac tatgccgatg ccaaaggcaa tgtgcaggct gatgatgcca tcagcaaccc
      781 caatggcagc atcaacagca ttgcgggtat tctgagcgaa aacaaacgca tttgcggcat
      841 gatgcctcat ccagaagatc tggttgatcc tctcatgggt ggggaagatg gtctgccatt
      901 gttccaaggt ctggtggagg ctgtagtccg gtgaataagc ctgtaaccgt tgatgaatcc
      961 cttgcccgtg aattcgagct ccgtcgacaa gcttgcggcc gcactcgagt ctggtaaaga
     1021 aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt
     1081 aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1141 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     1201 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1261 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1321 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1381 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1441 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1501 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1561 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1621 aattttaaca aaatattaac gtttacaatt tctggcggca cgatggcatg agattatcaa
     1681 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1741 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1801 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1861 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1921 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1981 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2041 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2101 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2161 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2221 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2281 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2341 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     2401 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     2461 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     2521 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2581 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2641 aatcatgatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2701 atttagaaaa ataaacaaat aggtcatgac caaaatccct taacgtgagt tttcgttcca
     2761 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2821 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2881 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2941 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3001 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3061 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3121 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3181 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3241 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3301 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3361 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3421 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3481 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3541 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     3601 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     3661 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     3721 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3781 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3841 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     3901 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     3961 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     4021 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     4081 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     4141 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     4201 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     4261 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     4321 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     4381 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     4441 tcctcaacga caggagcacg atcatgctag tcatgccccg cgcccaccgg aaggagctga
     4501 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     4561 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4621 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     4681 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     4741 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     4801 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     4861 atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     4921 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     4981 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     5041 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     5101 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     5161 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     5221 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     5281 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     5341 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     5401 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     5461 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     5521 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     5581 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     5641 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     5701 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     5761 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     5821 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     5881 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     5941 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     6001 ggccacgatg cgtccggcgt agaggatcga gatcgatctc gatcccgcga aattaatacg
     6061 actcactata
//
