LOCUS       sequence-431788-        8561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the ectodomain(1-797) of the short isoform of the Very
            Low-Density Lipoprotein Receptor (VLDLR, UniProtKB; P98155-2) fused 
            to a C-tag (EPEA) in th C- terminal for purification..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8561)
  TITLE     Hartl Lab plasmids
REFERENCE   2  (bases 1 to 8561)
  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"
COMMENT     Sequence Label: pB-T-PAF-VLDLR_ecto_short isoform
FEATURES             Location/Qualifiers
     source          1..8561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     583..605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     597..614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     621..640
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        621..639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   706..768
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1140..1158
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1177..1195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1214..1232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1251..1269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1288..1306
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1325..1343
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1362..1380
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1399..1417
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1439..1463
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(3969..3986)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3975..4199
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    4483..4530
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     rep_origin      4624..4758
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4763..5365
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     5262..5282
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5599..5720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5636..5655)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6188..6222
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(6371..6389)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6410..6426)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6410..6426)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6423..6445)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6434..6450
                     /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)."
     promoter        complement(join(6458..6464,6465..6482,6483..6488))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6503..6524
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6641..6658)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6812..7400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6892..6911)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7571..8362,8363..8431))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8194..8213
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8432..8536)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      601 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctcggtattc
      661 acgacagcag gctgaataat aaaaaaatta gaaactatta tttaacccta gaaagataat
      721 catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg ttttatcggt
      781 ctgtatatcg aggtttattt attaatttga atagatatta agttttatta tatttacact
      841 tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt attaaaaaaa
      901 aacaaaaact caaaatttct tctataaagt aacaaaactt ttaaacattc tctcttttac
      961 aaaaataaac ttattttgta ctttaaaaac agtcatgttg tattataaaa taagtaatta
     1021 gcttaaccta tacataatag aaacaaatta tacttattag tcagtcagaa acaactttgg
     1081 cacatatcaa tattatgctc tcgttaatta actcgaggga tccaagctgg ccgcacgtct
     1141 ccctatcagt gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta
     1201 cgtctagaac gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag
     1261 tgatagagaa ggtacgtcta gaacgtctcc ctatcagtga tagagaagtc gacacgtctc
     1321 gagctcccta tcagtgatag agaaggtacg tctagaacgt ctccctatca gtgatagaga
     1381 agtcgacacg tctcgagctc cctatcagtg atagagaagg taccccctat ataagcagag
     1441 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1501 gaagacaccg ggaccgatcc agcctccgcg gtggcggcca tcacaagttt gtacagatgg
     1561 gcacgtccgc gctctgggcg ctctggctgc tgctcgcgct gtgctgggcg ccccgggaga
     1621 gcggcgccac cggaaccggg agaaaagcca aatgtgaacc ctcccaattc cagtgcacaa
     1681 atggtcgctg tattacgctg ttgtggaaat gtgatgggga tgaagactgt gttgacggca
     1741 gtgatgaaaa gaactgtgta aagaagacgt gtgctgaatc tgacttcgtg tgcaacaatg
     1801 gccagtgtgt tcccagccga tggaagtgtg atggagatcc tgactgcgaa gatggttcag
     1861 atgaaagccc agaacagtgc catatgagaa catgccgcat acatgaaatc agctgtggcg
     1921 cccattctac tcagtgtatc ccagtgtcct ggagatgtga tggtgaaaat gattgtgaca
     1981 gtggagaaga tgaagaaaac tgtggcaata taacatgtag tcccgacgag ttcacctgct
     2041 ccagtggccg ctgcatctcc aggaactttg tatgcaatgg ccaggatgac tgcagcgatg
     2101 gcagtgatga gctggactgt gccccgccaa cctgtggcgc ccatgagttc cagtgcagca
     2161 cctcctcctg catccccatc agctgggtat gcgacgatga tgcagactgc tccgaccaat
     2221 ctgatgagtc cctggagcag tgtggccgtc agccagtcat acacaccaag tgtccagcca
     2281 gcgaaatcca gtgcggctct ggcgagtgca tccataagaa gtggcgatgt gatggggacc
     2341 ctgactgcaa ggatggcagt gatgaggtca actgtccctc tcgaacttgc cgacctgacc
     2401 aatttgaatg tgaggatggc agctgcatcc atggcagcag gcagtgtaat ggtatccgag
     2461 actgtgtcga tggttccgat gaagtcaact gcaaaaatgt caatcagtgc ttgggccctg
     2521 gaaaattcaa gtgcagaagt ggagaatgca tagatatcag caaagtatgt aaccaggagc
     2581 aggactgcag ggactggagt gatgagcccc tgaaagagtg tcatataaac gaatgcttgg
     2641 taaataatgg tggatgttct catatctgca aagacctagt tataggctac gagtgtgact
     2701 gtgcagctgg gtttgaactg atagatagga aaacctgtgg agatattgat gaatgccaaa
     2761 atccaggaat ctgcagtcaa atttgtatca acttaaaagg cggttacaag tgtgaatgta
     2821 gtcgtggcta tcaaatggat cttgctactg gcgtgtgcaa ggcagtaggc aaagagccaa
     2881 gtctgatctt cactaatcga agagacatca ggaagattgg cttagagagg aaagaatata
     2941 tccaactagt tgaacagcta agaaacactg tggctctcga tgctgacatt gctgcccaga
     3001 aactattctg ggccgatcta agccaaaagg ctatcttcag tgcctcaatt gatgacaagg
     3061 ttggtagaca tgttaaaatg atcgacaatg tctataatcc tgcagccatt gctgttgatt
     3121 gggtgtacaa gaccatctac tggactgatg cggcttctaa gactatttca gtagctaccc
     3181 tagatggaac caagaggaag ttcctgttta actctgactt gcgagagcct gcctccatag
     3241 ctgtggaccc actgtctggc tttgtttact ggtcagactg gggtgaacca gctaaaatag
     3301 aaaaagcagg aatgaatgga ttcgatagac gtccactggt gacagcggat atccagtggc
     3361 ctaacggaat tacacttgac cttataaaaa gtcgcctcta ttggcttgat tctaagttgc
     3421 acatgttatc cagcgtggac ttgaatggcc aagatcgtag gatagtacta aagtctctgg
     3481 agttcctagc tcatcctctt gcactaacaa tatttgagga tcgtgtctac tggatagatg
     3541 gggaaaatga agcagtctat ggtgccaata aattcactgg atcagagcta gccactctag
     3601 tcaacaacct gaatgatgcc caggacatca ttgtctatca tgaacttgta cagccatcag
     3661 gtaaaaattg gtgtgaagaa gacatggaga atggaggatg tgaataccta tgcctgccag
     3721 caccacagat taatgatcac tctccaaaat atacctgttc ctgtcccagt gggtacaatg
     3781 tagaggaaaa tggccgagac tgtcaaagga tcaatgtgac cacagcagta tcagaggtca
     3841 gtgttccccc aaaagggact tctggagaac cagaagccta agcatagata actgatccag
     3901 tgtgctgggc tacgatgcat gggcccccta ggtcgagtct agagggcccg tttaaacccg
     3961 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4021 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4081 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4141 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     4201 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4261 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4321 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4381 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4441 cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc tattccgaag
     4501 ttcctattct ctagaaagta taggaacttc cttggccaaa aagaccggta ccaggcaggc
     4561 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4621 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4681 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4741 ggaggctttt ttggaggcta ccatgggtac cgagtacaag cccacggtgc gcctcgccac
     4801 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4861 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4921 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4981 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5041 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5101 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5161 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5221 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5281 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5341 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag
     5401 gagcgcacga ccccatggct ccgaccgaag ccacccgggg cggccccgcc gaccccgcac
     5461 ccgcccccga ggcccaccga ctctagataa ctgatcataa tcagccatac cacatttgta
     5521 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     5581 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5641 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5701 aaactcatca atgtatctta acgctcgagt gcattctagt tgtggtttgt ccaaactcat
     5761 caatgtatct tatcatgtct gtataccgtc gacctctagc tgcaggaatt cgatatcaag
     5821 ctctagccaa caagctcgtc atcgctttgc agaagagcag agaggatatg ctcatcgtct
     5881 aaagaactac ccattttatt atatattagt cacgatatct ataacaagaa aatatatata
     5941 taataagtta tcacgtaagt agaacatgaa ataacaatat aattatcgta tgagttaaat
     6001 cttaaaagtc acgtaaaaga taatcatgcg tcattttgac tcacgcggtc gttatagttc
     6061 aaaatcagtg acacttaccg cattgacaag cacgcctcac gggagctcca agcggcgact
     6121 gagatgtcct aaatgcacag cgacggattc gcgctattta gaaagagaga gcaatatttc
     6181 aagaatgcat gcgtcaattt tacgcagact atctttctag ggttaaaaaa gatttgcgct
     6241 ttactcgacc taaactttaa acacgtcata gaatcttcgt ttgacaaaaa ccacattgtg
     6301 gccaagctgt gtgacgcgac gcgcgctaaa gaatggcaaa ccaagtcgcg cgaggtacca
     6361 gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt
     6421 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6481 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6541 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6601 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6661 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6721 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6781 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6841 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6901 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6961 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7021 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7081 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7141 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7201 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7261 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7321 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7381 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7441 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7501 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7561 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7621 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7681 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7741 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7801 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7861 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7921 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7981 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8041 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8101 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8161 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8221 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8281 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8341 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8401 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8461 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8521 aaataggggt tccgcgcaca tttccccgaa aagtgccacc t
//
