LOCUS       sequence-8249-ex        4676 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 4676)
  AUTHORS   Park D, Lapteva N, Seethammagari M, Slawin KM, Spencer DM
  TITLE     An essential role for Akt1 in dendritic cell function and tumor 
            immunotherapy.
  JOURNAL   Nat Biotechnol. 2006 Dec . 24(12):1581-90.
  PUBMED    17143278
REFERENCE   2  (bases 1 to 4676)
  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: "Nat 
            Biotechnol. 2006 Dec . 24(12):1581-90."
COMMENT     Sequence Label: pSH1/Mf-del-Akt-E
FEATURES             Location/Qualifiers
     source          1..4676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..83)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(317..336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(996..1787,1788..1856))
                     /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     1619..1638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1857..1961)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2029..2047
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2109..2129)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(2114..2443,2444..2459,2460..2729)
                     /label=SR-alpha promoter
                     /note="hybrid promoter consisting of the SV40 early 
                     promoter plus part of the long terminal repeat of human 
                     T-lymphotrophic virus 1"
     rep_origin      2294..2429
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     LTR             2460..2729
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          2774..2867
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     regulatory      2938..2947
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4147..4173
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4204..4338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4228..4247)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4282..4301
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4478..4500)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4641..4660
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
       61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1201 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1321 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1621 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1861 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1921 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1981 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2041 acgaggccct ttcgtcttca aggtcgatcg acctcatgtt tgacagctta tcatcgataa
     2101 gcttctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2161 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2221 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2281 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2341 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2401 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tccgatcgag
     2461 gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccaacgccgg
     2521 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag
     2581 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct
     2641 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt
     2701 tcgttttctg ttctgcgccg ttacagatcc aagctgcctc gatcgaggaa ctgaaaaacc
     2761 agaaagttaa ctggtaagtt tagtcttttt gtcttttatt tcaggtcccg gatccggtgg
     2821 tgcaaatcaa agaactgctc ctcagtggat gttgccttta cttctaggcc tgtacggagg
     2881 tgttacttct gctctaaaag ctgctctagc cctcgatcga gggctagagc ggccgcggcc
     2941 accatgggct gtgtgcaatg taaggataaa gaagcaacaa aactgacgga ggagctcgac
     3001 accgccattc agactgtggc cgatggactc aagaggcagg aagaagagac gatggacttc
     3061 cgatcaggct cacccagtga caactcaggg gctgaagaga tggaggtgtc cctggccaag
     3121 cccaagcacc gtgtgaccat gaacgagttt gagtacctga aactactggg caagggcacc
     3181 tttgggaaag tgattctggt gaaagagaag gccacaggcc gctactatgc catgaagatc
     3241 ctcaagaagg aggtcatcgt cgccaaggat gaggttgccc acacgcttac tgagaaccgt
     3301 gtcctgcaga actctaggca tcccttcctt acggccctca agtactcatt ccagacccac
     3361 gaccgcctct gctttgtcat ggagtatgcc aacgggggcg agctcttctt ccacctgtct
     3421 cgagagcgcg tgttctccga ggaccgggcc cgcttctatg gtgcggagat tgtgtctgcc
     3481 ctggactact tgcactccga gaagaacgtg gtgtaccggg acctgaagct ggagaacctc
     3541 atgctggaca aggacgggca catcaagata acggacttcg ggctgtgcaa ggaggggatc
     3601 aaggatggtg ccactatgaa gacattctgc ggaacgccgg agtacctggc ccctgaggtg
     3661 ctggaggaca acgactacgg ccgtgcagtg gactggtggg ggctgggcgt ggtcatgtat
     3721 gagatgatgt gtggccgcct gcccttctac aaccaggacc acgagaagct gttcgagctg
     3781 atcctcatgg aggagatccg cttcccgcgc acactcggcc ctgaggccaa gtccctgctc
     3841 tccgggctgc tcaagaagga ccctacacag aggctcggtg ggggctctga ggatgccaag
     3901 gagatcatgc agcaccggtt ctttgccaac atcgtgtggc aggatgtgta tgagaagaag
     3961 ctgagcccac ctttcaagcc ccaggtcacc tctgagactg acaccaggta tttcgatgag
     4021 gagttcacag ctcagatgat caccatcacg ccgcctgatc aagatgacag catggagtgt
     4081 gtggacagtg agcggaggcc gcacttcccc cagttctcct actcagccag tggcacagcc
     4141 gtcgactatc cgtacgacgt accagactac gcactcgact aagaattcct gattgattga
     4201 ccggatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4261 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4321 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4381 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     4441 gatcctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4501 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4561 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     4621 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accata
//
