LOCUS       .                       9385 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..1179
                     /label="EF-1α promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
     promoter        1196..1214
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1293..1317
                     /label="attB1"
                     /note="recombination site for the Gateway® BP reaction"
     CDS             1338..1358
                     /label="myr"
                     /note="N-myristoylation signal from Src kinase (Pellman et
                     al., 1985; Kaplan et al., 1988)"
     CDS             1371..2810
                     /label="Akt1"
                     /note="human serine/threonine protein kinase activated by
                     growth factors"
     CDS             2811..2837
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     protein_bind    complement(2857..2881)
                     /label="attB2"
                     /note="recombination site for the Gateway® BP reaction"
     CDS             2934..2975
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     misc_feature    2998..3574
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             3575..5224
                     /label="luciferase"
                     /note="firefly luciferase"
     CDS             5228..5269
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     polyA_signal    5313..5537
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    5607..5640
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(6098..6114)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6588..6692
                     /label="AmpR promoter"
     CDS             6693..7550
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      7724..8312
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    8600..8621
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8636..8666
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    8674..8690
                     /label="lac operator"
                     /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-β-D-thiogalactopyranoside (IPTG)."
     primer_bind     8698..8714
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        9387..10564
                     /label="EF-1α promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctccg
     1321 cggccgcccc cttcaccatg gggagcagca agagcaagcc caagtctaga atgaacgacg
     1381 tagccattgt gaaggagggc tggctgcaca aacgagggga atatattaaa acctggcggc
     1441 cacgctactt cctcctcaag aacgatggca cctttattgg ctacaaggaa cggcctcagg
     1501 atgtggatca gcgagagtcc ccactcaaca acttctcagt ggcacaatgc cagctgatga
     1561 agacagagcg gccaaggccc aacaccttta tcatccgctg cctgcagtgg accacagtca
     1621 ttgagcgcac cttccatgtg gaaacgcctg aggagcggga agaatgggcc accgccattc
     1681 agactgtggc cgatggactc aagaggcagg aagaagagac gatggacttc cgatcaggct
     1741 cacccagtga caactcaggg gctgaagaga tggaggtgtc cctggccaag cccaagcacc
     1801 gtgtgaccat gaacgagttt gagtacctga aactactggg caagggcacc tttgggaaag
     1861 tgattctggt gaaagagaag gccacaggcc gctactatgc catgaagatc ctcaagaagg
     1921 aggtcatcgt cgccaaggat gaggttgccc acacgcttac tgagaaccgt gtcctgcaga
     1981 actctaggca tcccttcctt acggccctca agtactcatt ccagacccac gaccgcctct
     2041 gctttgtcat ggagtatgcc aacgggggcg agctcttctt ccacctgtct cgagagcgcg
     2101 tgttctccga ggaccgggcc cgcttctatg gtgcggagat tgtgtctgcc ctggactact
     2161 tgcactccga gaagaacgtg gtgtaccggg acctgaagct ggagaacctc atgctggaca
     2221 aggacgggca catcaagata acggacttcg ggctgtgcaa ggaggggatc aaggatggtg
     2281 ccactatgaa gacattctgc ggaacgccgg agtacctggc ccctgaggtg ctggaggaca
     2341 acgactacgg ccgtgcagtg gactggtggg ggctgggcgt ggtcatgtat gagatgatgt
     2401 gtggccgcct gcccttctac aaccaggacc acgagaagct gttcgagctg atcctcatgg
     2461 aggagatccg cttcccgcgc acactcggcc ctgaggccaa gtccctgctc tccgggctgc
     2521 tcaagaagga ccctacacag aggctcggtg ggggctctga ggatgccaag gagatcatgc
     2581 agcaccggtt ctttgccaac atcgtgtggc aggatgtgta tgagaagaag ctgagcccac
     2641 ctttcaagcc ccaggtcacc tctgagactg acaccaggta tttcgatgag gagttcacag
     2701 ctcagatgat caccatcacg ccgcctgatc aagatgacag catggagtgt gtggacagtg
     2761 agcggaggcc gcacttcccc cagttctcct actcagccag tggcacagcc tacccatacg
     2821 acgtcccaga ctacgcttga aagggtgggc gcgccgaccc agctttcttg tacaaagtgg
     2881 ttgatatcca gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaggtaagc
     2941 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa tgagtttccc
     3001 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     3061 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3121 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3181 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     3241 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3301 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3361 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3421 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     3481 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3541 ctttgaaaaa cacgatgata atatggccac aaccatggaa gacgccaaaa acataaagaa
     3601 aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc
     3661 tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt
     3721 ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg
     3781 atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt
     3841 tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta
     3901 taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc
     3961 caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat
     4021 tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac
     4081 atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga
     4141 caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc
     4201 tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa
     4261 tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat
     4321 gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga
     4381 agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc
     4441 aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt
     4501 acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa
     4561 gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat
     4621 tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt
     4681 tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga
     4741 actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa
     4801 cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga
     4861 cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt
     4921 ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt
     4981 cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca
     5041 cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc
     5101 gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa
     5161 actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc
     5221 cgtgtaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtta
     5281 gtaatgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     5341 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     5401 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     5461 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     5521 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     5581 ccacgcgccc tgtagcggcg cattaaataa cttcgtataa tgtatgctat acgaagttat
     5641 atcgtatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
     5701 aactgacaca cattccacag aattccttga aatacatcca caggtacacc tccaattgac
     5761 tcaaatgatg tcaattagtc tatcagaagc ttctaaagcc atgacatcat tttctggaat
     5821 tttccaagct gtttaaaggc acagtcaact tagtgtatgt aaacttctga cccactggaa
     5881 ttgtgataca gtgaattata agtgaaataa tctgtctgta aacaattgtt ggaaaaatga
     5941 cttgtgtcat gcacaaagta gatgtcctaa ctgacttgcc aaaactattg tttgttaaca
     6001 agaaatttgt ggagtagttg aaaaacgagt tttaatgact ccaacttaag tgtatgtaaa
     6061 cttccgactt caactgtagg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc
     6121 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     6181 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     6241 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     6301 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     6361 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     6421 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     6481 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     6541 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     6601 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     6661 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6721 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6781 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6841 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6901 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6961 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     7021 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     7081 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     7141 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     7201 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     7261 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     7321 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7381 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     7441 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     7501 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     7561 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     7621 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     7681 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7741 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7801 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7861 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7921 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7981 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8041 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8101 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     8161 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     8221 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     8281 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     8341 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     8401 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     8461 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     8521 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     8581 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     8641 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     8701 gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac tctagaccta
     8761 tacagttgaa gtcggaagtt tacatacact taagttggag tcattaaaac tcgtttttca
     8821 actactccac aaatttcttg ttaacaaaca atagttttgg caagtcagtt aggacatcta
     8881 ctttgtgcat gacacaagtc atttttccaa caattgttta cagacagatt atttcactta
     8941 taattcactg tatcacaatt ccagtgggtc agaagtttac atacactaag ttgactgtgc
     9001 ctttaaacag cttggaaaat tccagaaaat gatgtcatgg ctttagaagc ttctgataga
     9061 ctaattgaca tcatttgagt caattggagg tgtacctgtg gatgtatttc aaggggatcc
     9121 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agcggccgct
     9181 agctcgagat cttaattaaa ctagtatcga tataacttcg tataatgtat gctatacgaa
     9241 gttatatgta cgggccagat atacgcgttg acattgatta ttgactaggc ttttgcaaaa
     9301 agctttgcaa agatggataa agttttaaac agagaggaat ctttgcagct aatggacctt
     9361 ctaggtcttg aaaggagtgg gaatt
//
