LOCUS       sequence-172954-        5634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of GST-CAR extracellular domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5634)
  AUTHORS   Patzke C, Max KE, Behlke J, Schreiber J, Schmidt H, Dorner AA, 
            Kroger S, Henning M, Otto A, Heinemann U, Rathjen FG
  TITLE     The coxsackievirus-adenovirus receptor reveals complex homophilic 
            and heterophilic interactions on neural cells.
  JOURNAL   J Neurosci. 2010 Feb 24;30(8):2897-910. doi: 
            10.1523/JNEUROSCI.5725-09.2010.
  PUBMED    20181587
REFERENCE   2  (bases 1 to 5634)
  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"; doi: 
            "10.1523/JNEUROSCI.5725-09"; journalName: "J Neurosci"; date: 
            "2010-02-24- 24"; volume: "30"; issue: "8"; pages: "2897-910"
COMMENT     Sequence Label: pGEX-chCARex
FEATURES             Location/Qualifiers
     source          1..5634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(530..535,536..553,554..560)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    568..584
                     /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     592..608
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     592..608
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             604..1131
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(624..641)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(624..640)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(633..655)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1233..1238,1239..1254,1255..1261)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1269..1285
                     /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             1308..1961
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1919..1941
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1968..1991
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2734..2756)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2901..2919)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2987..3091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3092..3160,3161..3952)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3310..3329)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4123..4711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4612..4631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4865..4882
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4955..5032
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5033..5634,1..481)
                     /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(5052..5071)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
       61 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
      121 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
      181 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
      241 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
      301 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
      361 ctcactggtg aaaagaaaaa ccacgctagc gcccaatacg caaaccgcct ctccccgcgc
      421 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      481 agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct ttacacttta
      541 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      601 gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
      661 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      721 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      781 cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt
      841 cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc
      901 atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat
      961 ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag
     1021 acgcgaatta tttttgatgg cgttggaatt agcttatcga ctgcacggtg caccaatgct
     1081 tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca
     1141 taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat
     1201 aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat
     1261 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agtattcatg tcccctatac
     1321 taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg
     1381 aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa
     1441 agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat
     1501 taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt
     1561 gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg
     1621 gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca
     1681 agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg
     1741 gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca
     1801 tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag
     1861 ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg
     1921 gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg gaagttctgt
     1981 tccaggggcc cctgggatcc ccggaattcc taagtataac ttcagctgaa tcagcatttg
     2041 aaaaagctca aggagagaga gttacgttgc catgtacctt tgaactctca gaagaggatg
     2101 taggcacact agatattgag tgggttttga taccagcaga tattcagaag aaggaagaaa
     2161 caataattct atattctgga gataggattt ataatcatta tcatcctgct ctggctgggc
     2221 ggctgcaatt tactagttct gatcccaaat ctggtgatgg ttcagtggat atcctgaatt
     2281 taaagtcagc agatactggc acataccagt gcaaagtgaa gaaggctcct ggagttgaaa
     2341 gcctaaaaat acagttgaac gtacttgtaa agccagcaag cactaaatgc tccattgaag
     2401 gatcacagga gattggaaag gacattgtat tgaagtgtgc atcacaagaa ggaaccccac
     2461 ttttgtatta tgactggaga agagtagtaa ctggcacaca aggacttcct gccacttccg
     2521 tactaaacaa aaatacaggg gaacttctct tgaaaaatgc ctctaaagac tattctggta
     2581 catacagttg tgttgcttca aaccgagttg gcacagatga atgttctgtt gagctgaatg
     2641 tcacccctcc tataaataca gctgtcgact cgagcggccg catcgtgact gactgacgat
     2701 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2761 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     2821 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     2881 taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     2941 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3001 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3061 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3121 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3181 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3241 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3301 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     3361 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3421 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3481 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3541 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3601 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     3661 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3721 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3781 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3841 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3901 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3961 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4021 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4081 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4141 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4201 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4261 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4321 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4381 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4441 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4501 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4561 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4621 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4681 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4741 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4801 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4861 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     4921 gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggc gcaaaacctt
     4981 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     5041 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     5101 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     5161 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     5221 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     5281 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     5341 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     5401 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     5461 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     5521 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     5581 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctg
//
