LOCUS       sequence-193769-        6549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged human LPL in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6549)
  AUTHORS   Chi X, Shetty SK, Shows HW, Hjelmaas AJ, Malcolm EK, Davies BS
  TITLE     Angiopoietin-like 4 Modifies the Interactions between Lipoprotein 
            Lipase and Its Endothelial Cell Transporter GPIHBP1.
  JOURNAL   J Biol Chem. 2015 May 8;290(19):11865-77. doi: 
            10.1074/jbc.M114.623769. Epub 2015 Mar 25.
  PUBMED    25809481
REFERENCE   2  (bases 1 to 6549)
  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.M114.623769"; journalName: "J Biol Chem"; date: 
            "2015-05-8- 8"; volume: "290"; issue: "19"; pages: "11865-77"
COMMENT     Sequence Label: pSS1 (hLPL-FLAG)
FEATURES             Location/Qualifiers
     source          1..6549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2432..2455
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2465..2482
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2505..2522)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2511..2735
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2781..3209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2868..2887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3078..3099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3218..3238)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3223..3552
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3403..3538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3465..3484
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3600..3647
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3666..4064
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4222..4343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4259..4278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4313..4332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4392..4408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4392..4408)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4405..4427)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4416..4432
                     /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(4440..4446,4447..4464,4465..4470))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4485..4506
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4623..4640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4794..5382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4874..4893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5553..6344,6345..6413))
                     /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     6176..6195
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6414..6518)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatggagag caaagccctg ctcgtgctga ctctggccgt
      961 gtggctccag agtctgaccg cctcccgcgg aggggtggcc gccgccgacc aaagaagaga
     1021 ttttatcgac atcgaaagta aatttgccct aaggacccct gaagacacag ctgaggacac
     1081 ttgccacctc attcccggag tagcagagtc cgtggctacc tgtcatttca atcacagcag
     1141 caaaaccttc atggtgatcc atggctggac ggtaacagga atgtatgaga gttgggtgcc
     1201 aaaacttgtg gccgccctgt acaagagaga accagactcc aatgtcattg tggtggactg
     1261 gctgtcacgg gctcaggagc attacccagt gtccgcgggc tacaccaaac tggtgggaca
     1321 ggatgtggcc cggtttatca actggatgga ggaggagttt aactaccctc tggacaatgt
     1381 ccatctcttg ggatacagcc ttggagccca tgctgctggc attgcaggaa gtctgaccaa
     1441 taagaaagtc aacagaatta ctggcctcga tccagctgga cctaactttg agtatgcaga
     1501 agccccgagt cgtctttctc ctgatgatgc agattttgta gacgtcttac acacattcac
     1561 cagagggtcc cctggtcgaa gcattggaat ccagaaacca gttgggcatg ttgacattta
     1621 cccgaatgga ggtacttttc agccaggatg taacattgga gaagctatcc gcgtgattgc
     1681 agagagagga cttggagatg tggaccagct agtgaagtgc tcccacgagc gctccattca
     1741 tctcttcatc gactctctgt tgaatgaaga aaatccaagt aaggcctaca ggtgcagttc
     1801 caaggaagcc tttgagaaag ggctctgctt gagttgtaga aagaaccgct gcaacaatct
     1861 gggctatgag atcaataaag tcagagccaa aagaagcagc aaaatgtacc tgaagactcg
     1921 ttctcagatg ccctacaaag tcttccatta ccaagtaaag attcattttt ctgggactga
     1981 gagtgaaacc cataccaatc aggcctttga gatttctctg tatggcaccg tggccgagag
     2041 tgagaacatc ccattcactc tgcctgaagt ttccacaaat aagacctact ccttcctaat
     2101 ttacacagag gtagatattg gagaactact catgttgaag ctcaaatgga agagtgattc
     2161 atactttagc tggtcagact ggtggagcag tcccggcttc gccattcaga agatcagagt
     2221 aaaagcagga gagactcaga aaaaggtgat cttctgttct agggagaaag tgtctcattt
     2281 gcagaaagga aaggcacctg cggtatttgt gaaatgccat gacaagtctc tgaataagaa
     2341 gtcaggcgga tccactagtc cagtgtggtg gaattctgca gatatccagc acagtggcgg
     2401 ccgctcgagt ctagagggcc cgcggttcga agattacaag gatgacgatg acaagcgtac
     2461 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     2521 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2581 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2641 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2701 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2761 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2821 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2881 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2941 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3001 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3061 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3121 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3181 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3241 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3301 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3361 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3421 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3481 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3541 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     3601 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     3661 aaaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     3721 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
     3781 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
     3841 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
     3901 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
     3961 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
     4021 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagcactt cgtggccgag
     4081 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4141 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4201 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4261 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4321 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4381 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4441 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4501 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4561 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4621 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4681 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4741 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4801 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4861 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4921 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4981 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5041 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5101 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5161 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5221 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5281 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5341 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5401 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5461 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5521 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5581 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5641 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5701 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5761 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5821 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5881 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5941 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6001 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6061 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6121 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6181 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6241 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6301 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6361 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6421 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6481 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6541 cctgacgtc
//
