LOCUS       sequence-327904-        6106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LDL-receptor associated protein (RAP), full length in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6106)
  AUTHORS   Hennen E, Safina D, Haussmann U, Worsdorfer P, Edenhofer F, Poetsch 
            A, Faissner A
  TITLE     A LewisX glycoprotein screen identifies the low density lipoprotein 
            receptor-related protein 1 (LRP1) as a modulator of 
            oligodendrogenesis in mice.
  JOURNAL   J Biol Chem. 2013 Jun 7;288(23):16538-45. doi: 
            10.1074/jbc.M112.419812. Epub 2013 Apr 24.
  PUBMED    23615909
REFERENCE   2  (bases 1 to 6106)
  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: 
            "10.1074/jbc.M112.419812"; journalName: "J Biol Chem"; date: 
            "2013-06-7- 7"; volume: "288"; issue: "23"; pages: "16538-45"
COMMENT     Sequence Label: RAPpGEX
FEATURES             Location/Qualifiers
     source          1..6106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      858..867
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1974..1996)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2141..2159)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2227..2331
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2332..2400,2401..3192)
                     /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(2550..2569)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3363..3951
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3852..3871
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4105..4122
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4195..4272
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4273..5355
                     /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(4292..4311)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5404..5409,5410..5427,5428..5434)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5442..5458
                     /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     5466..5482
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5466..5482
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5478..6005
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5498..5515)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5498..5514)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5507..5529)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctggt tccgcgtgga tccaggaaga tggcgcctcg aagagagagg
      781 gtctctacgc tgccccggct ccaactgcta gtgctgttgt tgctgccgct gatgcttgtg
      841 ccccagccca tagcaggcca tggcggcaag tactcgcgag agaagaacga gccggagatg
      901 gccgccaagc gcgagtccgg ggaggagttc cgcatggaga agctgaacca gctatgggag
      961 aaggccaagc ggcttcatct gtctcctgtg aggctggccg agctgcattc tgacctgaag
     1021 atacaagaga gggatgaact caactggaaa aagctgaagg tggaaggctt ggataaggat
     1081 ggggagaaag aagcaaaact gatccacaac ctcaacgtca tcctggccag atacggactg
     1141 gatgggagga aggacgccca gatggtgcac agcaacgccc tcaatgaaga cacccaggat
     1201 gagctggggg accccaggct ggaaaagctg tggcacaagg caaagacatc agggaaattc
     1261 tccagtgaag agctggacaa gctgtggaga gagtttctgc attacaaaga gaagatccag
     1321 gagtacaatg tgctgctaga cacactgagc agagctgaag aaggttatga gaaccttctc
     1381 agtccctcgg acatggccca catcaagagc gacaccctga tcagcaagca cagtgagctg
     1441 aaggacagac tgcgcagtat caaccagggc ttggaccgcc tgcggaaggt cagccaccag
     1501 ggctatggct ccaccactga gtttgaagag ccccgggtga tagatctgtg ggacctggct
     1561 cagtctgcca acttcactga gaaggaactg gagtcattca gggaggagct caagcacttt
     1621 gaggccaaaa ttgaaaagca caaccactac cagaagcagc tggagatttc ccaccagaag
     1681 ctgaagcacg tggagagcat cggcgacccc gagcacatca gccgcaacaa ggagaaatac
     1741 gtgctgctgg aggagaagac caaggagctg ggctacaagg tgaagaagca tctacaggac
     1801 ctgtctagca gggtctcaag ggctcggcac aatgagctct gaggaccaga agccaccagc
     1861 agcagcctag agagaacact tgaagacacc gggagctgtc agcatgtcct cgagcggccg
     1921 catcgtgact gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     1981 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     2041 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     2101 cgtagcgata gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt
     2161 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     2221 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2281 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2341 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2401 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2461 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2521 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     2581 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2641 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2701 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2761 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2821 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     2881 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2941 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3001 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3061 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3121 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3181 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3241 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3301 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3361 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3421 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3481 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3541 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3601 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3661 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3721 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3781 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3841 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3901 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3961 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4021 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4081 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     4141 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc
     4201 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     4261 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     4321 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     4381 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     4441 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     4501 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     4561 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     4621 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     4681 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     4741 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     4801 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     4861 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     4921 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     4981 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     5041 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     5101 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     5161 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     5221 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     5281 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5341 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca
     5401 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     5461 tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc
     5521 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     5581 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     5641 tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc
     5701 tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg
     5761 gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg
     5821 ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc
     5881 tacaggaagg ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac
     5941 ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt
     6001 cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt
     6061 tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctg
//
