LOCUS       sequence-211060-        6357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses StrepII-tagged mouse ANGPTL3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6357)
  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 6357)
  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: pHS18 (mANGPTL3-Strep)
FEATURES             Location/Qualifiers
     source          1..6357
                     /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             2319..2342
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2365..2382)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2371..2595
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2641..3069
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2728..2747)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2938..2959
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3078..3098)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3243..3377
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3304..3323
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3439..3486
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3505..3903
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4061..4182
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4098..4117)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4152..4171
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4231..4247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4231..4247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4244..4266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4255..4271
                     /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(4279..4285,4286..4303,4304..4309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4324..4345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4462..4479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4633..5221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4713..4732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5392..6183,6184..6252))
                     /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     6015..6034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6253..6357)
                     /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 accatgcaca
      961 caattaaatt attccttttt gttgttcctt tagtaattgc atccagagtg gatccagacc
     1021 tttcatcatt tgattctgca ccttcagagc caaaatcaag atttgctatg ttggatgatg
     1081 tcaaaatttt agcgaatggc ctcctgcagc tgggtcatgg acttaaagat tttgtccata
     1141 agactaaggg acaaattaac gacatatttc agaagctcaa catatttgat cagtcttttt
     1201 atgacctatc acttcgaacc aatgaaatca aagaagagga aaaggagcta agaagaacta
     1261 catctacact acaagttaaa aacgaggagg tgaagaacat gtcagtagaa ctgaactcaa
     1321 agcttgagag tctgctggaa gagaagacag cccttcaaca caaggtcagg gctttggagg
     1381 agcagctaac caacttaatt ctaagcccag ctggggctca ggagcaccca gaagtaacat
     1441 cactcaaaag ttttgtagaa cagcaagaca acagcataag agaactcctc cagagtgtgg
     1501 aagaacagta taaacaatta agtcaacagc acatgcagat aaaagaaata gaaaagcagc
     1561 tcagaaagac tggtattcaa gaaccctcag aaaattctct ttcttctaaa tcaagagcac
     1621 caagaactac tccccctctt caactgaacg aaacagaaaa tacagaacaa gatgaccttc
     1681 ctgccgactg ctctgccgtt tataacagag gcgaacatac aagtggcgtg tacactatta
     1741 aaccaagaaa ctcccaaggg tttaatgtct actgtgatac ccaatcaggc agtccatgga
     1801 cattaattca acaccggaaa gatggctcac aggacttcaa cgaaacatgg gaaaactacg
     1861 aaaagggctt tgggaggctc gatggagaat tttggttggg cctagagaag atctatgcta
     1921 tagtccaaca gtctaactac attttacgac tcgagctaca agactggaaa gacagcaagc
     1981 actacgttga atactccttt cacctgggca gtcacgaaac caactacacg ctacatgtgg
     2041 ctgagattgc tggcaatatc cccggggccc tcccagagca cacagacctg atgttttcta
     2101 catggaatca cagagcaaag ggacagctct actgtccaga aagttactca ggtggctggt
     2161 ggtggaatga catatgtgga gaaaacaacc taaatggaaa atacaacaaa cccagaacca
     2221 aatccagacc agagagaaga agagggatct actggagacc tcagagcaga aagctctatg
     2281 ctatcaaatc atccaaaatg atgctccagc ccaccacctg gagccacccg cagttcgaaa
     2341 aatgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2401 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2461 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2521 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2581 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2641 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2701 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2761 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2821 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2881 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2941 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3001 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3061 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3241 agtcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3301 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3361 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3421 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     3481 tacgacaagg tgaggaacta aaccatggcc aagcctttgt ctcaagaaga atccaccctc
     3541 attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc
     3601 agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact
     3661 gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac
     3721 ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg
     3781 tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat
     3841 ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc
     3901 taagcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
     3961 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4021 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4081 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4141 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4201 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4261 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4321 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4381 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4441 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4501 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4561 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4621 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4681 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4741 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4801 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4861 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4921 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4981 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5041 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5101 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5161 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5221 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5281 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5341 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5401 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5461 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5521 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5581 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5641 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5701 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5761 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5821 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5881 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5941 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6001 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6061 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6121 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6181 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6241 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6301 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcg
//
