LOCUS       62056_20520        7064 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7064)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     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(R) BP reaction"
     CDS             1338..1358
                     /codon_start=1
                     /label=myr
                     /note="N-myristoylation signal from Src kinase (Pellman et
                     al., 1985; Kaplan et al., 1988)"
                     /translation="MGSSKSK"
     CDS             1371..2810
                     /codon_start=1
                     /label=Akt1
                     /note="human serine/threonine protein kinase activated by
                     growth factors"
                     /translation="MNDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQRESPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWATAIQ
                     TVADGLKRQEEETMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK
                     VILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR
                     LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLM
                     LDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMY
                     EMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPTQRLGGGSEDA
                     KEIMQHRFFANIVWQDVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSM
                     ECVDSERRPHFPQFSYSASGTA"
     protein_bind    complement(2830..2854)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2907..2948
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    2992..3216
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3286..3319
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3777..3793)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4267..4371
                     /label=AmpR promoter
     CDS             4372..5229
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5403..5991
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6279..6300
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6315..6345
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6353..6369
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6377..6393
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7064
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
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 tgaaagggtg
     2821 ggcgcgccga cccagctttc ttgtacaaag tggttgatat ccagcacagt ggcggccgct
     2881 cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg
     2941 attctacgcg taccggttag taatgagttt aaacccgctg atcagcctcg actgtgcctt
     3001 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3061 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3121 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3181 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3241 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaaataac ttcgtataat
     3301 gtatgctata cgaagttata tcgtatgcat gctttgcata cttctgcctg ctggggagcc
     3361 tggggacttt ccacacccta actgacacac attccacaga attccttgaa atacatccac
     3421 aggtacacct ccaattgact caaatgatgt caattagtct atcagaagct tctaaagcca
     3481 tgacatcatt ttctggaatt ttccaagctg tttaaaggca cagtcaactt agtgtatgta
     3541 aacttctgac ccactggaat tgtgatacag tgaattataa gtgaaataat ctgtctgtaa
     3601 acaattgttg gaaaaatgac ttgtgtcatg cacaaagtag atgtcctaac tgacttgcca
     3661 aaactattgt ttgttaacaa gaaatttgtg gagtagttga aaaacgagtt ttaatgactc
     3721 caacttaagt gtatgtaaac ttccgacttc aactgtaggg taccgagctc gaattcactg
     3781 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     3841 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     3901 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     3961 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     4021 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4081 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4141 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     4201 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4261 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4321 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4381 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4441 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4501 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4561 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4621 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4681 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4741 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4801 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4861 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4921 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4981 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5041 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5101 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5161 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5221 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5281 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5341 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5401 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5461 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5521 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5581 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5641 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5701 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5761 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5821 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5881 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5941 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6001 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6061 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6121 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6181 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     6241 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     6301 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     6361 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcctg
     6421 caggtcgact ctagacctat acagttgaag tcggaagttt acatacactt aagttggagt
     6481 cattaaaact cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc
     6541 aagtcagtta ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac
     6601 agacagatta tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca
     6661 tacactaagt tgactgtgcc tttaaacagc ttggaaaatt ccagaaaatg atgtcatggc
     6721 tttagaagct tctgatagac taattgacat catttgagtc aattggaggt gtacctgtgg
     6781 atgtatttca aggggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     6841 ctagaatgca gcggccgcta gctcgagatc ttaattaaac tagtatcgat ataacttcgt
     6901 ataatgtatg ctatacgaag ttatatgtac gggccagata tacgcgttga cattgattat
     6961 tgactaggct tttgcaaaaa gctttgcaaa gatggataaa gttttaaaca gagaggaatc
     7021 tttgcagcta atggaccttc taggtcttga aaggagtggg aatt
//
