LOCUS       sequence-318916-        7061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C. elegans iPGM containing a C-terminal biotinylation site
            and His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7061)
  AUTHORS   Wiedmann M, Dranchak PK, Aitha M, Queme B, Collmus CD, Kashipathy 
            MM, Kanter L, Lamy L, Rogers JM, Tao D, Battaile KP, Rai G, Lovell 
            S, Suga H, Inglese J
  TITLE     Structure-activity relationship of ipglycermide binding to 
            phosphoglycerate mutases.
  JOURNAL   J Biol Chem. 2021 Apr 1:100628. doi: 10.1016/j.jbc.2021.100628.
  PUBMED    33812994
REFERENCE   2  (bases 1 to 7061)
  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: "J Biol 
            Chem. 2021 Apr 1:100628. doi: 10.1016/j.jbc.2021.100628."
COMMENT     Sequence Label: pET21a Ce-iPGM-bio
FEATURES             Location/Qualifiers
     source          1..7061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             89..121
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             1700..1744
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1745..1762
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1793..1810
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1863..1881)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1877..1924
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1961..2416
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2048..2067)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2258..2279
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2442..2546
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2547..2615,2616..3407)
                     /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     complement(2765..2784)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3578..4166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4067..4086
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4320..4337
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4352..4491
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4417..4436)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4577..4599
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4593..4784)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5593..6675)
                     /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     6637..6656
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6676..6753)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6959..6978
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggctagcatg actggtggac agcaaatggg
      121 tcgcggatcc atggcgatgg caaataacag ttcggtggcc aataaggtct gtctcatcgt
      181 tattgatgga tggggagttt ctgaagatcc ttacggtaac gctattctca acgcacagac
      241 accagttatg gacaagctgt gttcgggcaa ttgggctcaa attgaggcac atggtcttca
      301 tgttggtctc ccagaaggat tgatgggaaa ttcggaagtc ggacatttga acatcggagc
      361 cggacgtgtt atctatcaag acattgttcg tattaatctg gcagtcaaga acaacaaatt
      421 tgtgactaat gagagcttgg tggatgcttg cgatcgtgct aaaaacggaa atggacgtct
      481 tcatctggcc ggacttgttt ctgacggagg tgttcattct catattgatc acatgtttgc
      541 tttggttaag gccatcaaag agctcggagt tccagaactt taccttcatt tctacggaga
      601 tggtcgtgat acttctccaa acagtggagt tggattcctt gaacaaaccc tcgagttctt
      661 ggagaaaact actggatatg gaaaactagc tactgtagtt ggccgctact atgctatgga
      721 tcgcgataac agatgggagc gtatcaatgt tgcatacgag gcaatgattg gaggtgttgg
      781 agagacttcc gatgaggctg gggttgttga agttgttcgc aagcgttacg ctgctgatga
      841 aacagacgaa ttcttgaagc caatcattct tcaaggagag aaaggacgtg ttcaaaatga
      901 cgatacaatc atcttcttcg actaccgtgc tgatcgtatg cgtgagattt ctgcagcaat
      961 gggaatggat cgttacaagg attgcaattc gaagttagct catccatcaa atcttcaagt
     1021 atatggaatg actcaataca aagccgagtt cccattcaaa tcgctgttcc cgccagcatc
     1081 gaacaaaaat gtattggctg agtggctcgc cgagcaaaaa gtttcgcaat ttcattgtgc
     1141 ggaaaccgaa aaatacgctc acgttacatt tttcttcaat ggaggacttg aaaaacaatt
     1201 tgagggagaa gaaaggtgtt tagtgcccag tccaaaggtc gcaacttacg atcttcaacc
     1261 agaaatgtct gcggccggcg ttgctgacaa aatgattgaa caactcgagg ctggaactca
     1321 tccattcatt atgtgcaact ttgctccacc agatatggtc gggcatacgg gagtctatga
     1381 agctgctgtc aaggcctgtg aagctactga tatcgcaatc ggaagaatct atgaagcaac
     1441 tcaaaagcac ggatactcac ttatggttac tgctgatcac ggaaatgctg aaaagatgaa
     1501 ggctccagat ggtggaaaac acactgctca cacatgttac cgtgttccac tcactttgag
     1561 ccatccagga ttcaaatttg tcgatccagc cgaccgtcat ccggcccttt gtgatgttgc
     1621 tccaacagtt ctcgctatta tgggactccc tcaaccagct gaaatgactg gggtctcgat
     1681 tgttcagaag atcaagcttg gtctgaacga catcttcgag gctcagaaaa tcgaatggca
     1741 cgaattagtg cctcgcggaa gcgctgcagg gaagcttgcg gccgcactcg agcaccacca
     1801 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1861 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1921 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     1981 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2041 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2101 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2161 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2221 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2281 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2341 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2401 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2461 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2521 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2581 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2641 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2701 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2761 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2821 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2881 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2941 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3001 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3061 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     3121 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3181 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3241 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3301 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3361 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3421 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3481 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3541 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3601 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3661 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3721 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3781 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3841 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3901 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3961 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4021 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4081 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4141 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4201 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4261 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4321 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4381 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     4441 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4501 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4561 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4621 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4681 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     4741 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     4801 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     4861 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4921 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4981 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     5041 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     5101 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     5161 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     5221 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     5281 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     5341 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     5401 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     5461 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     5521 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     5581 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5641 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     5701 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     5761 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     5821 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     5881 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     5941 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     6001 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     6061 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     6121 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     6181 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     6241 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     6301 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     6361 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     6421 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     6481 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     6541 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     6601 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     6661 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     6721 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     6781 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     6841 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     6901 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     6961 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     7021 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa t
//
