LOCUS       sequence-211073-        6372 bp    DNA     circular SYN 14-APR-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 6372)
  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 6372)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..6372
                     /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"
     CDS             2334..2357
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2380..2397)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2386..2610
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2656..3084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2743..2762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2953..2974
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3093..3113)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3258..3392
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3319..3338
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3454..3501
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3520..3918
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4076..4197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4113..4132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4167..4186
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4246..4262)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4246..4262)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4259..4281)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4270..4286
                     /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(4294..4300,4301..4318,4319..4324))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4339..4360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4477..4494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4648..5236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4728..4747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5407..6198,6199..6267))
                     /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     6030..6049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6268..6372)
                     /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 accatgttca
      961 caattaagct ccttcttttt attgttcctc tagttatttc ctccagaatt gatcaagaca
     1021 attcatcatt tgattctcta tctccagagc caaaatcaag atttgctatg ttagacgatg
     1081 taaaaatttt agccaatggc ctccttcagt tgggacatgg tcttaaagac tttgtccata
     1141 agacgaaggg ccaaattaat gacatatttc aaaaactcaa catatttgat cagtcttttt
     1201 atgatctatc gctgcaaacc agtgaaatca aagaagaaga aaaggaactg agaagaacta
     1261 catataaact acaagtcaaa aatgaagagg taaagaatat gtcacttgaa ctcaactcaa
     1321 aacttgaaag cctcctagaa gaaaaaattc tacttcaaca aaaagtgaaa tatttagaag
     1381 agcaactaac taacttaatt caaaatcaac ctgaaactcc agaacaccca gaagtaactt
     1441 cacttaaaac ttttgtagaa aaacaagata atagcatcaa agaccttctc cagaccgtgg
     1501 aagaccaata taaacaatta aaccaacagc atagtcaaat aaaagaaata gaaaatcagc
     1561 tcagaaggac tagtattcaa gaacccacag aaatttctct atcttccaag ccaagagcac
     1621 caagaactac tccctttctt cagttgaatg aaataagaaa tgtaaaacat gatggcattc
     1681 ctgctgaatg taccaccatt tataacagag gtgaacatac aagtggcatg tatgccatca
     1741 gacccagcaa ctctcaagtt tttcatgtct actgtgatgt tatatcaggt agtccatgga
     1801 cattaattca acatcgaata gatggatcac aaaacttcaa tgaaacgtgg gagaactaca
     1861 aatatggttt tgggaggctt gatggagaat tttggttggg cctagagaag atatactcca
     1921 tagtgaagca atctaattat gttttacgaa ttgagttgga agactggaaa gacaacaaac
     1981 attatattga atattctttt tacttgggaa atcacgaaac caactatacg ctacatctag
     2041 ttgcgattac tggcaatgtc cccaatgcaa tcccggaaaa caaagatttg gtgttttcta
     2101 cttgggatca caaagcaaaa ggacacttca actgtccaga gggttattca ggaggctggt
     2161 ggtggcatga tgagtgtgga gaaaacaacc taaatggtaa atataacaaa ccaagagcaa
     2221 aatctaagcc agagaggaga agaggattat cttggaagtc tcaaaatgga aggttatact
     2281 ctataaaatc aaccaaaatg ttgatccatc caacagattc agaaagcttt gaatggagcc
     2341 acccgcagtt cgaaaaatga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     2401 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2461 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2521 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2581 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     2641 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2701 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2761 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2821 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2881 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2941 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3001 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3061 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3121 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3181 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3241 ttagtcagca accatagtcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3301 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3361 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3421 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
     3481 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagcc tttgtctcaa
     3541 gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa
     3601 gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat
     3661 gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct
     3721 gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg
     3781 agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata
     3841 gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt
     3901 tatgtgtggg agggctaagc acttcgtggc cgaggagcag gactgacacg tgctacgaga
     3961 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4021 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4081 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4141 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4201 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4261 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4321 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4381 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4441 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4501 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4561 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4621 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4681 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4741 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4801 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4861 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4921 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4981 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5041 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5101 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5161 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5221 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5281 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5341 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5401 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5461 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5521 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5581 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5641 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5701 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5761 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5821 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5881 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5941 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6001 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6061 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6121 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6181 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6241 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6301 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6361 aggggttccg cg
//
