LOCUS       sequence-366771-        6101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the human protein APOA4_HUMAN. This 
            is one of the bottom performing targets in our apoptosis assay (it 
            induced apoptosis)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6101)
  AUTHORS   Veling MT, Nguyen DT, Thadani NN, Oster ME, Rollins NJ, Brock KP, 
            Bethel NP, Lim S, Baker D, Way JC, Marks DS, Chang RL, Silver PA
  TITLE     Natural and Designed Proteins Inspired by Extremotolerant Organisms 
            Can Form Condensates and Attenuate Apoptosis in Human Cells.
  JOURNAL   ACS Synth Biol. 2022 Mar 18;11(3):1292-1302. doi: 
            10.1021/acssynbio.1c00572. Epub 2022 Feb 18.
  PUBMED    35176859
REFERENCE   2  (bases 1 to 6101)
  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: 
            "10.1021/acssynbio.1c00572"; journalName: "ACS Synth Biol"; date: 
            "2022-03-18- 18"; volume: "11"; issue: "3"; pages: "1292-1302"
COMMENT     Sequence Label: pST_APOA4_HUMAN (pBS0645)
FEATURES             Location/Qualifiers
     source          1..6101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2111..2128)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2117..2341
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2387..2815
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2474..2493)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2684..2705
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2824..2844)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2829..3158
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3009..3144
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3071..3090
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3206..3253
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3272..3646
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3776..3897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3813..3832)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3867..3886
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3946..3962)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3946..3962)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3959..3981)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3970..3986
                     /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(3994..4000,4001..4018,4019..4024))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4177..4194)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4348..4936)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4428..4447)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5107..5898,5899..5967))
                     /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     5730..5749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5968..6072)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gccaccatgt ttctgaaggc tgtggtgctg accctcgcat tggtggctgt ggctggggcc
      961 agagccgaag tgtcagcaga tcaggtggcc actgtgatgt gggattattt ttctcagctt
     1021 agtaacaacg ctaaagaagc agtggagcat ctgcagaaga gcgagctgac ccagcagtta
     1081 aacgccctgt tccaggacaa gctgggagag gttaacactt atgccggtga tctgcagaaa
     1141 aaactggtgc ctttcgccac cgagctgcac gaacgcctgg ccaaggattc cgaaaagctt
     1201 aaggaggaaa ttggcaagga gctcgaggag ctgcgggccc gattgctgcc acacgccaac
     1261 gaggtgtctc agaagatcgg cgacaatctt cgcgagctgc agcagagact cgagccctac
     1321 gccgaccagc tgagaaccca ggtcaacaca caggccgagc agctcagacg gcagctgacc
     1381 ccttatgctc agcggatgga gcgggtgctt agggagaacg ccgacagcct gcaagcctcc
     1441 ctccgccccc acgctgatga actcaaggca aagatcgacc agaacgtcga ggaactgaaa
     1501 ggcagactaa caccctacgc cgatgaattc aaagtgaaga ttgatcagac cgtagaggag
     1561 ctccggcggt ccttagcacc atatgcccag gatacccagg aaaagctcaa tcatcagctg
     1621 gagggcctga ctttccaaat gaaaaaaaac gctgaagaac tgaaggccag gatcagcgcc
     1681 agtgctgagg agctgagaca gagactggcc ccactcgccg aggacgtgcg cggcaacctc
     1741 cggggaaata cagaggggct gcagaagagc ctggctgagc tgggagggca cctggaccag
     1801 caggtggaag agttccggag aagggtggaa ccttatggcg agaattttaa caaagccctc
     1861 gtgcagcaga tggagcagct gcggcagaag ctagggcccc atgccggaga cgtggagggt
     1921 cacctgtctt ttctggagaa ggacctgaga gacaaggtga attcattctt ctctacattc
     1981 aaggaaaaag agagccagga taagaccctg tctctgcctg agctggagca gcaacaggag
     2041 cagcagcagg agcagcaaca ggagcaggtg cagatgctgg cccctctgga gagctgaacc
     2101 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2161 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2221 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2281 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2341 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2401 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2461 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2521 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2581 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2641 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2701 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2761 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2821 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2881 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3001 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3061 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3121 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3181 ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat
     3241 agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt
     3301 gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc
     3361 ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat
     3421 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg
     3481 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc
     3541 cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga
     3601 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga
     3661 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     3721 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     3781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3841 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3901 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     3961 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4021 taaagcctgg ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     4081 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4141 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4201 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4261 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4321 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4381 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4441 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4501 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4561 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4621 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4681 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4741 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4801 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4861 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4921 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4981 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5041 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5101 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5161 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5221 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5281 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5341 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5401 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5461 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5521 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5581 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5641 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5701 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5761 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5821 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5881 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5941 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6001 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6061 aggggttccg cgcacatttc cccgaaaagt gccacctgac g
//
