LOCUS       sequence-493864-        4683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4683)
  AUTHORS   Geleziunas R, Xu W, Takeda K, Ichijo H, Greene WC
  TITLE     HIV-1 Nef inhibits ASK1-dependent death signalling providing a 
            potential mechanism for protecting the infected host cell.
  JOURNAL   Nature. 2001 Apr 12. 410(6830):834-8.
  PUBMED    11298454
REFERENCE   2  (bases 1 to 4683)
  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"; journalName: "Nature. 
            2001 Apr 12. 410(6830):834-8."
COMMENT     Sequence Label: pCI NL4-3 Nef-HA-WT
FEATURES             Location/Qualifiers
     source          1..4683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        146..525
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        526..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          865..997
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     980..999
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1042..1060
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1729..1755
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1735..1761
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     polyA_signal    1805..1926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1816..1835)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1870..1889
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2107..2562
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2194..2213)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2404..2425
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2629..2648)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2748..2770
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2808..2826)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2894..2998
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2999..3067,3068..3859)
                     /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     complement(3217..3236)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4030..4618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4519..4538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acagatcttc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat
       61 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
      121 tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca
      181 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      241 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      301 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      361 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac
      421 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt
      481 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      541 cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      601 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      661 aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcaga
      841 agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt ttaaggagac
      901 caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt
      961 ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca gttcaattac
     1021 agctcttaag gctagagtac ttaatacgac tcactatagg ctagcctcga gaattagctt
     1081 gacagggctt ggaaaggatt ttgctataag atgggtggca agtggtcaaa aagtagtgtg
     1141 attggatggc ctgctgtaag ggaaagaatg agacgagctg agccagcagc agatggggtg
     1201 ggagcagtat ctcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct
     1261 aacaatgctg cttgtgcctg gctagaagca caagaggagg aagaggtggg ttttccagtc
     1321 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     1381 ttaaaagaaa aggggggact ggaagggcta attcactccc aaagaagaca agatatcctt
     1441 gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta cacaccaggg
     1501 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca
     1561 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg
     1621 catggaatgg atgaccctga gagagaagtg ttagagtgga ggtttgacag ccgcctagca
     1681 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgcta tccgtacgac
     1741 gtgcctgatt atgcatcatt gtgaatctag agtcgacccg ggcggccgct tcgagcagac
     1801 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1861 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1921 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     1981 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgggctgg
     2041 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2101 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2161 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2221 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2281 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2341 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2401 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     2461 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     2521 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     2581 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     2641 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     2701 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2761 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     2821 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     2881 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2941 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3001 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3061 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3121 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3181 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3241 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3301 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3361 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3421 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3481 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3541 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3601 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3661 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3721 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3781 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3841 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3901 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3961 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4021 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4081 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4141 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4201 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4261 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4321 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4381 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4441 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4501 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4561 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4621 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatggc
     4681 tcg
//
