LOCUS       sequence-372573-        6143 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses hyrdoxyornithine transformylase PvdF from Pseudomonas 
            aeruginosa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6143)
  AUTHORS   Kenjic N, Hoag MR, Moraski GC, Caperelli CA, Moran GR, Lamb AL
  TITLE     PvdF of pyoverdin biosynthesis is a structurally unique 
            N(10)-formyltetrahydrofolate-dependent formyltransferase.
  JOURNAL   Arch Biochem Biophys. 2019 Mar 30;664:40-50. doi: 
            10.1016/j.abb.2019.01.028. Epub 2019 Jan 26.
  PUBMED    30689984
REFERENCE   2  (bases 1 to 6143)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Arch 
            Biochem Biophys."; date: "2019-03-30"; volume: "664:40-50. doi"; 
            pages: " 10.1016/j.abb.2019.01.028. Epub 2019 Jan 26"
COMMENT     Sequence Label: pET29b-pvdF WT
FEATURES             Location/Qualifiers
     source          1..6143
                     /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(215..232)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     RBS             1080..1102
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1117..1141
                     /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(1141..1160)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1142..1160)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1248..1267)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1473..1550
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1551..2633
                     /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(1570..1589)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3442..3633
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3627..3649)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3735..3874
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3790..3809
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3889..3906)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4060..4648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4140..4159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4759..4778)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4770..5585
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4770..5585
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4843..4862)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5677..6132)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5814..5835)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6026..6045
                     /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 cgaattcgga tccgatatcg ccatggaacc gcgtggcacc agggtaccca
      241 gtcatcagag cttctcggcg agcagcgcca gcccttcgtg caaggccggg aacaggctgt
      301 tgttgaagtt gttccagcgc agctcgagga tggtgtcgtc gggtgaaatc tccgttttca
      361 gcacatcatg gaacacttcg ccggaatcga tgccattgtc cacatagtgg aacgatgctc
      421 cggtccagta cagcggctcg accgcgacct tttccatggt cgcccaatcc accaccttct
      481 cgccccgcgc tccatacaac gcgtccaggg tcgcataggc gccacgacgc tcgtaaggcg
      541 agtcctcgcg cgtcacgcca ggatggatat tcatgatccg ccgtgcgaac ggagcgcccg
      601 ggcgtaccag ctcatcgagg atgaccagga gcccgtccaa taccaccaca tcggcgccca
      661 gctccaccag ggtgtcatgc aggcgtcgct cgaaatcgct cttgccggcc acatgctcgg
      721 gggtaccccg cgggtaccga cggtaggtgg acggcacgct gagcaagagg tcgttcaggg
      781 tcttgccttg cacctgcagg tctgccggat agaaccattg ctgcccggga cgataggcga
      841 aaccgtagtc cttgattttt ccctggtcgc gaggcagctc cgcgtcgtcg tcgtagatca
      901 cgcccacgag ctcgtaggca tcgcccagcg cagtctggtt cagcgcctcc accaaggact
      961 ccagtaccga cttcatgtac cgctgttcgc ccttgtacgg cacgtactgg ccggccttgt
     1021 cggccgcggc gtttctcagg gaccagatat aggccagttt ccttttcgtc atatgtatat
     1081 ctccttctta aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc
     1141 ccctatagtg agtcgtatta atttcgcggg atcgagatcg atctcgatcc tctacgccgg
     1201 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
     1261 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
     1321 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
     1381 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
     1441 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc
     1501 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     1561 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     1621 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     1681 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     1741 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     1801 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     1861 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     1921 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     1981 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     2041 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     2101 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     2161 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     2221 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     2281 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     2341 ttggtgcgga catctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     2401 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     2461 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     2521 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     2581 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     2641 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
     2701 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     2761 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     2821 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     2881 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     2941 gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg
     3001 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     3061 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     3121 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     3181 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     3241 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     3301 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     3361 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     3421 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     3481 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     3541 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
     3601 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3661 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3721 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     3781 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccattgcggt gtgaaatacc
     3841 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     3901 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4501 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg
     4741 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct
     4801 ctaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     4861 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
     4921 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     4981 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     5041 ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc caggtattag
     5101 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     5161 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     5221 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     5281 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     5341 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     5401 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     5461 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     5521 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     5581 tctaagaatt aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5641 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     5701 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     5761 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     5821 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     5881 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     5941 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     6001 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     6061 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     6121 tacagggcgc gtcccattcg cca
//
