LOCUS       sequence-8148-ex        7677 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 7677)
  AUTHORS   Scheid MP, Marignani PA, Woodgett JR
  TITLE     Multiple phosphoinositide 3-kinase-dependent steps in activation of 
            protein kinase B.
  JOURNAL   Mol Cell Biol. 2002 Sep . 22(17):6247-60.
  PUBMED    12167717
REFERENCE   2  (bases 1 to 7677)
  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: "Mol Cell 
            Biol. 2002 Sep . 22(17):6247-60."
COMMENT     Sequence Label: HA PKB T308D S473D pcDNA3
FEATURES             Location/Qualifiers
     source          1..7677
                     /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             962..988
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             968..994
                     /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"
     promoter        complement(3229..3247)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3230..3247)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3267..3284)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3273..3497
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3543..3971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3630..3649)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3840..3861
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3980..4000)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3985..4315
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4166..4301
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4228..4247
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4382..5176
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4436..4455)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5046..5065
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5350..5471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5387..5406)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5441..5460
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5520..5536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5520..5536)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5533..5555)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5544..5560
                     /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(5568..5574,5575..5592,5593..5598))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5613..5634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5751..5768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5922..6510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6002..6021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6681..7472,7473..7541))
                     /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     7304..7323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7542..7646)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcggatcct agaccaccat
      961 gtacccatac gatgttccag attacgcaag cttgggtggt cccatggacg acgtggccat
     1021 cgtgaaggag ggctggctgc acaagcgagg tgagtacatc aagacgtggc ggccgcgtta
     1081 cttcctcctg aagaacgatg gcacgttcat tggctacaag gagcggccgc aggacctgga
     1141 gcagcgtgag tcgcccctca acaacttctc cgtggcccaa tgccagctga tgaagacgga
     1201 gcggccgcgg cccaacacct tcatcatccg ctgcctgcag tggaccacgg tcatcgagcg
     1261 cacgttccac gtggagacgc ccgaggagcg ggaggagtgg accaccgcca tccagacggt
     1321 ggccgacggg ctcaagaggc aggaggagga gacgatggac ttccggtcgg gctcacccgg
     1381 cgagaactcg ggggccgagg agatggaggt gtcgctggcc aagcccaagc accgcgtgac
     1441 catgaatgag tttgagtacg tgaagctgct gggcaaaggc actttcggga aggtgatcct
     1501 ggtgaaggag aaggccacag cggcctacta cgccatgaag atcctaaaga aggaggtcat
     1561 cgtggccaag gacgaggtgg cccacacgct cacagagaac cgcgttctcc agaactcccg
     1621 gcacccgtcc ctgacggccc tgaagtactc cttccagaca cacgaccgcc tgtgcttcgt
     1681 catggagtac gctaacgggg gcgagctttt cttccacctg tcccgggagc gggtgttttc
     1741 cgaggaccgg gcccgtttct acggcgccga gatcgtgtcg gccctggatt acctgcactc
     1801 ggaaaaggaa gtggtgtaca gggacctcaa gctggagaac ctcatgctgg acaaggacgg
     1861 gcacatcaag atcaccgact tcggactgtg caaggagggc atcaaggatg gcgccaccat
     1921 gaaggatttc tgcggtaccc ccgaatacct ggctcccgag gtgctggagg acaacgacta
     1981 cggccgggca gtggactggt gggggctggg tgtggtcatg tacgagatga tgtgcggccg
     2041 cctgcccttc tacaaccagg accacgagaa gctctttgag ctcattctca tggaggagat
     2101 ccgcttcccc cgcacactca gcccggaggc caagtccctg ctctctgggc tactcaagaa
     2161 ggaccccaag cagcggcttg gtgggggctc tgaggacgcc aaggagatca tgcagcaccg
     2221 attcttcgcc agcatcgtgt ggcaggacgt gtacgagaag aagctcagcc cgcccttcaa
     2281 gcctcaggtc acatctgaga cggacaccag gtattttgat gaggagttca cggcccagat
     2341 gatcaccatt acgccacctg accaagacga cagcatggag ggggtggaca gcgaacggag
     2401 gccccacttc ccccagttcg actactcggc tagcgcgacg gcctgagggg cccgctgccg
     2461 gagctcagta caccgccacg ggcagccacg tgctctggac gcttccgagg actgggagga
     2521 agcctcgcac ggaggatctg tatttaatgt tttttatttc tggatgcgtt tgagtggaac
     2581 ctcgccatcc tccaacacag gatggagagg accatggcgg ggcgaccgag aaccacctcc
     2641 tttgcggaga gctgcaagcc agtgcagcag ccttcagctt ttggtagcat gaaagttagc
     2701 agagataaag atggcagcaa ggtaaccaca gtggtggcaa ctcctggaca gggtcctgac
     2761 aggccacagg aagtcagtta cacagacact aaagtcattg gaaatgggtc atttggtgtg
     2821 gtatatcaag ccaaactttg tgactcagga gaactggtgg ccatcaagaa agttcttcag
     2881 gacaagcgat ttaagaaccg agagctccag atcatgagaa agctagatca ctgtaacata
     2941 gtccgattgc ggtatttctt ctactcgagt ggcgagaaga aagatgaggt ctaccttaac
     3001 ctggtgctgg actatgttcc ggaaacagtg tacagagtcg ccagacacta tagtcgagcc
     3061 aagcagacac tccctgtgat ctatgtcaag ttgtatatgt accagctgtt cagaagtcta
     3121 gcctatatcc attcctttgg gatctgccat cgagacatta aaccacagaa cctcttgctg
     3181 gatcgatcca gatcctctag agtcgagcat gcatctagag ggccctattc tatagtgtca
     3241 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3301 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3361 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3421 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3481 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3541 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3601 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3661 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     3721 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3781 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3841 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3901 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3961 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4021 ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4081 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4141 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4201 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4261 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4321 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4381 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4441 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4501 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4561 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4621 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4681 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4741 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4801 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4861 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4921 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4981 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5041 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5101 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5161 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5221 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5281 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5341 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5401 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5461 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5521 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5581 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5641 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5701 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5761 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5821 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5881 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5941 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6001 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6061 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     6121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6181 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6361 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6601 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6661 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6721 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6781 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6841 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6901 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6961 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7021 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7081 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7141 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7201 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7261 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7321 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7381 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7441 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7501 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7561 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7621 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
