LOCUS       sequence-11472-e        7855 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 7855)
  AUTHORS   Jones RG, Parsons M, Bonnard M, Chan VS, Yeh WC, Woodgett JR, Ohashi
            PS
  TITLE     Protein kinase B regulates T lymphocyte survival, nuclear factor 
            kappaB activation, and Bcl-X(L) levels in vivo.
  JOURNAL   J Exp Med. 2000 May 15. 191(10):1721-34.
  PUBMED    10811865
REFERENCE   2  (bases 1 to 7855)
  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: "J Exp Med. 
            2000 May 15. 191(10):1721-34."
COMMENT     Sequence Label: GAG-PKB-pcDNA3
FEATURES             Location/Qualifiers
     source          1..7855
                     /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..1378
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1308..1330
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1346..1362
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        complement(3407..3425)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3408..3425)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3445..3462)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3451..3675
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3721..4149
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3808..3827)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4018..4039
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4158..4178)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4163..4493
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4344..4479
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4406..4425
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4560..5354
                     /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(4614..4633)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5224..5243
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5528..5649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5565..5584)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5619..5638
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5698..5714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5698..5714)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5711..5733)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5722..5738
                     /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(5746..5752,5753..5770,5771..5776))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5791..5812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5929..5946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6100..6688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6180..6199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6859..7650,7651..7719))
                     /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     7482..7501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7720..7824)
                     /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 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     1021 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     1081 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     1141 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     1201 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt
     1261 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc
     1321 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccaa
     1381 acctaaacct caagttcttt ctgacagtgg ggggccgctc atcgacctac ttacagaaga
     1441 ccccccgcct tatagggacc caagaccacc cccttccgac agggacggaa atggtggaga
     1501 agcgacccct gcgggagagg caccggaccc ctccccaatg gcatctcgcc tacgtgggag
     1561 acgggagccc cctgtggccg actccactac ctcgcaggca ttccccctcc gcgcaggagg
     1621 aaacggacag cttcaatact ggccgttctc ctcttctgac ctttacaact ggaaaaataa
     1681 taacccttct ttttctgaag atccaggtaa actgacagct ctgatcgagt ctgttctcat
     1741 ctacccatac ccatggacga cgtggccatc gtgaaggagg gctggctgca caagcgaggt
     1801 gagtacatca agacgtggcg gccgcgttac ttcctcctga agaacgatgg cacgttcatt
     1861 ggctacaagg agcggccgca ggacctggag cagcgtgagt cgcccctcaa caacttctcc
     1921 gtggcccaat gccagctgat gaagacggag cggccgcggc ccaacacctt catcatccgc
     1981 tgcctgcagt ggaccacggt catcgagcgc acgttccacg tggagacgcc cgaggagcgg
     2041 gaggagtgga ccaccgccat ccagacggtg gccgacgggc tcaagaggca ggaggaggag
     2101 acgatggact tccggtcggg ctcacccggc gagaactcgg gggccgagga gatggaggtg
     2161 tcgctggcca agcccaagca ccgcgtgacc atgaatgagt ttgagtacgt gaagctgctg
     2221 ggcaaaggca ctttcgggaa ggtgatcctg gtgaaggaga aggccacagc ggcctactac
     2281 gccatgaaga tcctaaagaa ggaggtcatc gtggccaagg acgaggtggc ccacacgctc
     2341 acagagaacc gcgttctcca gaactcccgg cacccgtccc tgacggccct gaagtactcc
     2401 ttccagacac acgaccgcct gtgcttcgtc atggagtacg ctaacggggg cgagcttttc
     2461 ttccacctgt cccgggagcg ggtgttttcc gaggaccggg cccgtttcta cggcgccgag
     2521 atcgtgtcgg ccctggatta cctgcactcg gaaaaggaag tggtgtacag ggacctcaag
     2581 ctggagaacc tcatgctgga caaggacggg cacatcaaga tcaccgactt cggactgtgc
     2641 aaggagggca tcaaggatgg cgccaccatg aagactttct gcgggacccc cgaatacctg
     2701 gctcccgagg tgctggagga caacgactac ggccgggcag tggactggtg ggggctgggt
     2761 gtggtcatgt acgagatgat gtgcggccgc ctgcccttct acaaccagga ccacgagaag
     2821 ctctttgagc tcattctcat ggaggagatc cgcttccccc gcacactcag cccggaggcc
     2881 aagtccctgc tctctgggct actcaagaag gaccccaagc agcggcttgg tgggggctct
     2941 gaggacgcca aggagatcat gcagcaccga ttcttcgcca gcatcgtgtg gcaggacgtg
     3001 tacgagaaga agctcagccc gcccttcaag cctcaggtca catctgagac ggacaccagg
     3061 tattttgatg aggagttcac ggcccagatg atcaccatta cgccacctga ccaagacgac
     3121 agcatggagg gggtggacag cgaacggagg ccccacttcc cccagttctc ctactcggcc
     3181 agcgcgacgg cctgaggggc ccgctgccgg agctcagtac accgccacgg gcagccacgt
     3241 gctctggacg cttccgagga ctgggaggaa gcctcgcacg gaggatctgt atttaatgtt
     3301 ttttatttct ggatgcgttt gagtggaacc tcgccatcct ccaacacagg atggagagga
     3361 ccatggggga tcctctagag tcgagcatgc atctagaggg ccctattcta tagtgtcacc
     3421 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3481 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3541 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3601 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3661 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3721 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3781 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3841 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     3901 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3961 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4021 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4081 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4141 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4201 aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4261 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4321 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4381 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4441 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4501 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4561 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4621 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4681 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4741 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4801 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4861 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4921 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4981 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5041 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5101 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5161 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5221 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5281 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5341 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5401 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5461 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5521 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5581 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5641 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5701 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5761 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5821 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5881 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5941 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6001 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6061 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6241 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     6301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6421 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6481 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6541 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6601 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6661 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6721 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6781 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6841 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6901 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6961 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7021 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7081 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7141 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7201 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7261 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7321 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7381 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7441 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7501 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7561 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7621 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7681 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7741 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7801 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
