LOCUS       sequence-211083-        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(75..94)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        266..645
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        646..849
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     800..820
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     894..913
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        894..912
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      948..957
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2463..2486
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2496..2513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2536..2553)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2542..2766
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2812..3240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2899..2918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3109..3130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3249..3269)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3254..3583
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3434..3569
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3496..3515
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3631..3678
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3697..4095
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4253..4374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4290..4309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4344..4363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4423..4439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4423..4439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4436..4458)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4447..4463
                     /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(4471..4477,4478..4495,4496..4501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4516..4537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4654..4671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4825..5413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4905..4924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5584..6375,6376..6444))
                     /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     6207..6226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6445..6549)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
       61 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
      121 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
      181 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
      241 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
      301 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      361 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      421 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      481 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      541 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      601 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      661 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      721 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      781 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      841 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
      901 actcactata gggagaccca agctggctag cgtttaaact taagcttgcc accatggaga
      961 gcaaagccct gctcgtgctg actctggccg tgtggctcca gagtctgacc gcctcccgcg
     1021 gaggggtggc cgccgccgac caaagaagag attttatcga catcgaaagt aaatttgccc
     1081 taaggacccc tgaagacaca gctgaggaca cttgccacct cattcccgga gtagcagagt
     1141 ccgtggctac ctgtcatttc aatcacagca gcaaaacctt catggtgatc catggctgga
     1201 cggtaacagg aatgtatgag agttgggtgc caaaacttgt ggccgccctg tacaagagag
     1261 aaccagactc caatgtcatt gtggtggact ggctgtcacg ggctcaggag cattacccag
     1321 tgtccgcggg ctacaccaaa ctggtgggac aggatgtggc ccggtttatc aactggatgg
     1381 aggaggagtt taactaccct ctggacaatg tccatctctt gggatacagc cttggagccc
     1441 atgctgctgg cattgcagga agtctgacca ataagaaagt caacagaatt actggcctcg
     1501 atccagctgg acctaacttt gagtatgcag aagccccgag tcgtctttct cctgatgatg
     1561 cagattttgt agacgtctta cacacattca ccagagggtc ccctggtcga agcattggaa
     1621 tccagaaacc agttgggcat gttgacattt acccgaatgg aggtactttt cagccaggat
     1681 gtaacattgg agaagctatc cgcgtgattg cagagagagg acttggagat gtggaccagc
     1741 tagtgaagtg ctcccacgag cgctccattc atctcttcat cgactctctg ttgaatgaag
     1801 aaaatccaag taaggcctac aggtgcagtt ccaaggaagc ctttgagaaa gggctctgct
     1861 tgagttgtag aaagaaccgc tgcaacaatc tgggctatga gatcaataaa gtcagagcca
     1921 aaagaagcag caaaatgtac ctgaagactc gttctcagat gccctacaaa gtcttccatt
     1981 accaagtaaa gattcatttt tctgggactg agagtgaaac ccataccaat caggcctttg
     2041 agatttctct gtatggcacc gtggccgaga gtgagaacat cccattcact ctgcctgaag
     2101 tttccacaaa taagacctac tccttcctaa tttacacaga ggtagatatt ggagaactac
     2161 tcatgttgaa gctcaaatgg aagagtgatt catactttag ctggtcagac tggtggagca
     2221 gtcccggctt cgccattcag aagatcagag taaaagcagg agagactcag aaaaaggtga
     2281 tcttctgttc tagggagaaa gtgtctcatt tgcagaaagg aaaggcacct gcggtatttg
     2341 tgaaatgcca tgacaagtct ctgaataaga agtcaggcgg atccactagt ccagtgtggt
     2401 ggaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc ccgcggttcg
     2461 aagattacaa ggatgacgat gacaagcgta ccggtcatca tcaccatcac cattgagttt
     2521 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2581 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2641 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2701 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2761 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2821 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2881 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2941 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3001 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3061 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3121 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3181 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3241 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3301 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3361 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3421 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3481 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     3541 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     3601 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     3661 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa
     3721 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     3781 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     3841 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     3901 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     3961 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
     4021 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
     4081 gtgtgggagg gctaagcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt
     4141 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4201 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4261 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4321 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4381 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4441 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4501 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4561 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4621 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4681 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4741 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4801 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4861 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4921 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4981 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5041 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5101 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5161 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5221 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5281 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5341 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5401 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5461 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5521 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5581 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5641 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5701 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5761 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5821 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5881 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5941 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6001 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6061 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6121 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6181 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6241 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6301 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6361 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6421 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6481 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6541 ggttccgcg
//
