LOCUS       sequence-206091-        5652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V5 and His-tagged human ANGPTL8 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5652)
  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 5652)
  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: pEB30 (hANGPTL8-V5-His)
FEATURES             Location/Qualifiers
     source          1..5652
                     /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             1517..1558
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1568..1585
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1608..1625)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1614..1838
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1884..2312
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1971..1990)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2181..2202
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2321..2341)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2326..2655
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2506..2641
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2568..2587
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2703..2750
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2769..3167
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3325..3446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3362..3381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3416..3435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3495..3511)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3495..3511)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3508..3530)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3519..3535
                     /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(3543..3549,3550..3567,3568..3573))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3588..3609
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3726..3743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3897..4485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3977..3996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4656..5447,5448..5516))
                     /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     5279..5298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5517..5621)
                     /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 ccatgccagt gcctgctctg tgcctgctct gggccctggc
      961 aatggtgacc cggcctgcct cagcggcccc catgggcggc ccagaactgg cacagcatga
     1021 ggagctgacc ctgctcttcc atgggaccct gcagctgggc caggccctca acggtgtgta
     1081 caggaccacg gagggacggc tgacaaaggc caggaacagc ctgggtctct atggccgcac
     1141 aatagaactc ctggggcagg aggtcagccg gggccgggat gcagcccagg aacttcgggc
     1201 aagcctgttg gagactcaga tggaggagga tattctgcag ctgcaggcag aggccacagc
     1261 tgaggtgctg ggggaggtgg cccaggcaca gaaggtgcta cgggacagcg tgcagcggct
     1321 agaagtccag ctgaggagcg cctggctggg ccctgcctac cgagaatttg aggtcttaaa
     1381 ggctcacgct gacaagcaga gccacatcct atgggccctc acaggccacg tgcagcggca
     1441 gaggcgggag atggtggcac agcagcatcg gctgcgacag atccaggaga gactccacac
     1501 agcggctctc ccagccggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     1561 taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     1621 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1681 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1741 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1801 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     1861 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1921 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1981 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2041 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2101 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2161 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     2221 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     2281 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     2341 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2401 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2461 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2521 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2581 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2641 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     2701 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     2761 actaaaccat ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg
     2821 ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta
     2881 gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag
     2941 aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg
     3001 cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc
     3061 tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag
     3121 ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaagca cttcgtggcc
     3181 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3241 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3301 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     3361 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3421 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     3481 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3541 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3601 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3661 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3721 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3781 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3841 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3901 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3961 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4021 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4081 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4141 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4201 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4261 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4321 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4381 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4441 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4501 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4561 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4621 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4681 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4741 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4801 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4861 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4921 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4981 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5041 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5101 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5161 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5221 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5281 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5341 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5401 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5461 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5521 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5581 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5641 ccacctgacg tc
//
