LOCUS       sequence-478573-        6695 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Gorilla ACOD1 in E.coli expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6695)
  AUTHORS   Chen F, Yalcin I, Zhao M, Chen C, Blankenfeldt W, Pessler F, Bussow 
            K
  TITLE     Amino acid positions near the active site determine the reduced 
            activity of human ACOD1 compared to murine ACOD1.
  JOURNAL   Sci Rep. 2023 Jun 26;13(1):10360. doi: 10.1038/s41598-023-37373-w.
  PUBMED    37365251
REFERENCE   2  (bases 1 to 6695)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2023-06-26"; volume: "13(1):10360. doi"; pages: " 
            10.1038/s41598-023-37373-w"
COMMENT     Sequence Label: pCAD183_gCAD_pET
FEATURES             Location/Qualifiers
     source          1..6695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     206..225
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     309..328
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        309..327
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    328..352
                     /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             367..389
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             405..428
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             435..455
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1867..1884
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1937..1955)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1951..1998
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2035..2490
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2122..2141)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2332..2353
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2583..3398)
                     /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             complement(2583..3398)
                     /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     3306..3325
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3390..3409
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3520..4108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4009..4028
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4262..4279
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4294..4433
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4359..4378)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4519..4541
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4535..4726)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5535..6617)
                     /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     6579..6598
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6618..6695)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
       61 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
      121 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
      181 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
      241 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
      301 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
      361 aataattttg tttaacttta agaaggagat atacaatggc tagctggagc cacccgcagt
      421 tcgaaaaagt cgacgagaac ctctacttcc agggaggcgg ccgcaagtct atcacagaaa
      481 gctttgccac agcaatccat ggcttgaaag tgggacacct gacagatcgt gttattcaga
      541 ggagcaagag gatgattcta gacactctgg gtgctgggtt cctgggaacc actacggaag
      601 tgtttcacat agccagccaa tatagcaaga tctacagttc caacatatcc agcactgttt
      661 ggggtcagcc agacatcagg ctcccgccca catatgctgc ttttgtgaac ggtgtggcta
      721 ttcactccat ggattttgat gacacgtggc accctgccac ccacccttct ggggctgtcc
      781 ttcctgtcct cacagcttta gcagaagccc tgccaaggag tccaaagttt tctggccttg
      841 acctgctgct ggctttcaat gttggtattg aagtgcaagg ccgattactg catttcgcca
      901 aggaggccaa tgacatacca aagagattcc atcccccttc cgtggtagga acgttgggta
      961 gtgctgctgc tgcatccaag tttttaggac ttagctcgac aaagtgccga gaagctctgg
     1021 ccattgctgt ttcccatgct ggggcaccca tggccaatgc tgccacccag accaagcccc
     1081 tccacattgg caatgctgcc aagcatggga tagaagctgc atttttggca atgttgggtc
     1141 tgcaaggaaa caagcaggtc ttggacttgg aggcaggatt tggggccttt tatgccaact
     1201 attccccaaa agtccttcca agcctggctt cctacagttg gctgctggac cagcaggacg
     1261 tggcctttaa gcgttttcct gcacatttat ctacccactg ggtggcagac gcagctgcat
     1321 ctgtgagaaa gcaccttgta gcagagagag ccctgcttcc aactgactac attaagagaa
     1381 ttgtgctcag gataccaaat gtccagtatg taaacaggcc ctttccagtt tcggagcatg
     1441 aagcccgtca ttcattccag tatgtggcct gtgccatgct gcttgatggt ggcatcactg
     1501 tcccctcatt ccatgaatgc cagatcaaca ggccacaggt gagagagctg ctcagtaagg
     1561 tggagctgga gtaccctccg gacaacttgc caagcttcaa caccctgtac tgtgaaataa
     1621 gtgtcaccct caaggatgga gccaccttca cagatcgctc tgataccttc tatgggcact
     1681 ggagaaaacc actgagccag gaggacctag aggaaaagtt cagagccaat gcctccaaga
     1741 tgctgtcctg ggacacagtg gaaagcctta taaagatagt cgaaaatcta gaagacctag
     1801 aagactgttc tgtgttaact atacttctca aaggaccctg aaggttagta agcggccgca
     1861 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     1921 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     1981 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
     2041 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2101 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2161 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2221 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2281 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2341 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2401 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2461 attttaacaa actagtaacg tttacaattt caggtggcac ttttcgggga aatgtgcgcg
     2521 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     2581 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     2641 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2701 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2761 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2821 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     2881 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     2941 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     3001 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tgttttcacc tgaatcagga
     3061 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     3121 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     3181 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     3241 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     3301 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     3361 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     3421 atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca
     3481 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3541 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3601 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3661 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3721 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3781 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3841 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3901 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3961 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4021 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4081 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4141 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4201 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4261 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     4321 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     4381 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     4441 acccgccaac accagctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4501 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4561 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     4621 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     4681 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     4741 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     4801 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     4861 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     4921 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     4981 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     5041 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     5101 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     5161 tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct
     5221 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     5281 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     5341 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     5401 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     5461 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     5521 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5581 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     5641 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     5701 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     5761 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     5821 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     5881 ttggcaacca gcatcgcagt gggaacgata ccctcattca gcatttgcat ggtttgttga
     5941 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     6001 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     6061 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     6121 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     6181 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     6241 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     6301 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     6361 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     6421 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     6481 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     6541 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     6601 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     6661 ataccgcgaa aggttttgcg ccattcgatg gtgtc
//
