LOCUS       sequence-206245-        6393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses StrepII-tagged human ANGPTL3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6393)
  AUTHORS   Chi X, Britt EC, Shows HW, Hjelmaas AJ, Shetty SK, Cushing EM, Li W,
            Dou A, Zhang R, Davies BSJ
  TITLE     ANGPTL8 promotes the ability of ANGPTL3 to bind and inhibit 
            lipoprotein lipase.
  JOURNAL   Mol Metab. 2017 Oct;6(10):1137-1149. doi: 
            10.1016/j.molmet.2017.06.014. Epub 2017 Jun 29.
  PUBMED    29031715
REFERENCE   2  (bases 1 to 6393)
  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: 
            "10.1016/j.molmet.2017.06.014"; journalName: "Mol Metab"; date: 
            "2017-10"; volume: "6"; issue: "10"; pages: "1137-1149"
COMMENT     Sequence Label: pHS15 (hANGPTL3-Strep)
FEATURES             Location/Qualifiers
     source          1..6393
                     /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"
     CDS             2303..2326
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2349..2366)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2355..2579
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2625..3053
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2712..2731)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2922..2943
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3062..3082)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3067..3396
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3247..3382
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3309..3328
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3444..3491
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3510..3908
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4066..4187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4103..4122)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4157..4176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4236..4252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4236..4252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4249..4271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4260..4276
                     /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(4284..4290,4291..4308,4309..4314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4329..4350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4467..4484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4638..5226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4718..4737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5397..6188,6189..6257))
                     /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     6020..6039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6258..6362)
                     /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 ccatgttcac aattaagctc cttcttttta ttgttcctct
      961 agttatttcc tccagaattg atcaagacaa ttcatcattt gattctctat ctccagagcc
     1021 aaaatcaaga tttgctatgt tagacgatgt aaaaatttta gccaatggcc tccttcagtt
     1081 gggacatggt cttaaagact ttgtccataa gacgaagggc caaattaatg acatatttca
     1141 aaaactcaac atatttgatc agtcttttta tgatctatcg ctgcaaacca gtgaaatcaa
     1201 agaagaagaa aaggaactga gaagaactac atataaacta caagtcaaaa atgaagaggt
     1261 aaagaatatg tcacttgaac tcaactcaaa acttgaaagc ctcctagaag aaaaaattct
     1321 acttcaacaa aaagtgaaat atttagaaga gcaactaact aacttaattc aaaatcaacc
     1381 tgaaactcca gaacacccag aagtaacttc acttaaaact tttgtagaaa aacaagataa
     1441 tagcatcaaa gaccttctcc agaccgtgga agaccaatat aaacaattaa accaacagca
     1501 tagtcaaata aaagaaatag aaaatcagct cagaaggact agtattcaag aacccacaga
     1561 aatttctcta tcttccaagc caagagcacc aagaactact ccctttcttc agttgaatga
     1621 aataagaaat gtaaaacatg atggcattcc tgctgaatgt accaccattt ataacagagg
     1681 tgaacataca agtggcatgt atgccatcag acccagcaac tctcaagttt ttcatgtcta
     1741 ctgtgatgtt atatcaggta gtccatggac attaattcaa catcgaatag atggatcaca
     1801 aaacttcaat gaaacgtggg agaactacaa atatggtttt gggaggcttg atggagaatt
     1861 ttggttgggc ctagagaaga tatactccat agtgaagcaa tctaattatg ttttacgaat
     1921 tgagttggaa gactggaaag acaacaaaca ttatattgaa tattcttttt acttgggaaa
     1981 tcacgaaacc aactatacgc tacatctagt tgcgattact ggcaatgtcc ccaatgcaat
     2041 cccggaaaac aaagatttgg tgttttctac ttgggatcac aaagcaaaag gacacttcaa
     2101 ctgtccagag ggttattcag gaggctggtg gtggcatgat gagtgtggag aaaacaacct
     2161 aaatggtaaa tataacaaac caagagcaaa atctaagcca gagaggagaa gaggattatc
     2221 ttggaagtct caaaatggaa ggttatactc tataaaatca accaaaatgt tgatccatcc
     2281 aacagattca gaaagctttg aatggagcca cccgcagttc gaaaaatgag tttaaacccg
     2341 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2401 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2461 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2521 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     2581 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     2641 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2701 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2761 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2821 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2881 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2941 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3001 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3061 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3121 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3181 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3241 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3301 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3361 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3421 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
     3481 tataatacga caaggtgagg aactaaacca tggccaagcc tttgtctcaa gaagaatcca
     3541 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
     3601 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
     3661 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
     3721 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
     3781 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
     3841 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
     3901 agggctaagc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
     3961 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4021 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4081 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4141 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4201 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4261 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4321 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4381 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4441 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4501 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4561 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4621 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4681 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4741 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4801 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4861 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4921 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4981 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5041 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5101 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5161 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5221 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5281 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5341 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5401 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5461 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5521 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5581 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5641 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5701 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5761 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5821 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5881 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5941 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6001 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6061 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6121 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6181 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6241 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6301 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6361 cgcacatttc cccgaaaagt gccacctgac gtc
//
