LOCUS       62056_20525        7940 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 7940)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7940
                     /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             1341..1364
                     /codon_start=1
                     /label=OVA peptide
                     /note="antigenic CD8+ T cell epitope from chicken
                     ovalbumin"
                     /translation="SIINFEKL"
     CDS             1437..2129
                     /codon_start=1
                     /label=ZsGreen1
                     /note="Zoanthus green fluorescent protein"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     CDS             2187..2207
                     /codon_start=1
                     /label=myr
                     /note="N-myristoylation signal from Src kinase (Pellman et
                     al., 1985; Kaplan et al., 1988)"
                     /translation="MGSSKSK"
     CDS             2220..3659
                     /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"
     CDS             3660..3686
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     protein_bind    complement(3706..3730)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3783..3824
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3868..4092
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    4162..4195
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4653..4669)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5143..5247
                     /label=AmpR promoter
     CDS             5248..6105
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6279..6867
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7155..7176
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7191..7221
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7229..7245
                     /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     7253..7269
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7940
                     /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 agtataatca actttgaaaa actgggttct ggcgagggca
     1381 gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggcccttcc ggaaccatgg
     1441 cccagtccaa gcacggcctg accaaggaga tgaccatgaa gtaccgcatg gagggctgcg
     1501 tggacggcca caagttcgtg atcaccggcg agggcatcgg ctaccccttc aagggcaagc
     1561 aggccatcaa cctgtgcgtg gtggagggcg gccccttgcc cttcgccgag gacatcttgt
     1621 ccgccgcctt catgtacggc aaccgcgtgt tcaccgagta cccccaggac atcgtcgact
     1681 acttcaagaa ctcctgcccc gccggctaca cctgggaccg ctccttcctg ttcgaggacg
     1741 gcgccgtgtg catctgcaac gccgacatca ccgtgagcgt ggaggagaac tgcatgtacc
     1801 acgagtccaa gttctacggc gtgaacttcc ccgccgacgg ccccgtgatg aagaagatga
     1861 ccgacaactg ggagccctcc tgcgagaaga tcatccccgt gcccaagcag ggcatcttga
     1921 agggcgacgt gagcatgtac ctgctgctga aggacggtgg ccgcttgcgc tgccagttcg
     1981 acaccgtgta caaggccaag tccgtgcccc gcaagatgcc cgactggcac ttcatccagc
     2041 acaagctgac ccgcgaggac cgcagcgacg ccaagaacca gaagtggcac ctgaccgagc
     2101 acgccatcgc ctccggctcc gccttgcccg caacaaactt ctctctgctg aaacaagccg
     2161 gagatgtcga agagaatcct ggaccgatgg ggagcagcaa gagcaagccc aagtctagaa
     2221 tgaacgacgt agccattgtg aaggagggct ggctgcacaa acgaggggaa tatattaaaa
     2281 cctggcggcc acgctacttc ctcctcaaga acgatggcac ctttattggc tacaaggaac
     2341 ggcctcagga tgtggatcag cgagagtccc cactcaacaa cttctcagtg gcacaatgcc
     2401 agctgatgaa gacagagcgg ccaaggccca acacctttat catccgctgc ctgcagtgga
     2461 ccacagtcat tgagcgcacc ttccatgtgg aaacgcctga ggagcgggaa gaatgggcca
     2521 ccgccattca gactgtggcc gatggactca agaggcagga agaagagacg atggacttcc
     2581 gatcaggctc acccagtgac aactcagggg ctgaagagat ggaggtgtcc ctggccaagc
     2641 ccaagcaccg tgtgaccatg aacgagtttg agtacctgaa actactgggc aagggcacct
     2701 ttgggaaagt gattctggtg aaagagaagg ccacaggccg ctactatgcc atgaagatcc
     2761 tcaagaagga ggtcatcgtc gccaaggatg aggttgccca cacgcttact gagaaccgtg
     2821 tcctgcagaa ctctaggcat cccttcctta cggccctcaa gtactcattc cagacccacg
     2881 accgcctctg ctttgtcatg gagtatgcca acgggggcga gctcttcttc cacctgtctc
     2941 gagagcgcgt gttctccgag gaccgggccc gcttctatgg tgcggagatt gtgtctgccc
     3001 tggactactt gcactccgag aagaacgtgg tgtaccggga cctgaagctg gagaacctca
     3061 tgctggacaa ggacgggcac atcaagataa cggacttcgg gctgtgcaag gaggggatca
     3121 aggatggtgc cactatgaag acattctgcg gaacgccgga gtacctggcc cctgaggtgc
     3181 tggaggacaa cgactacggc cgtgcagtgg actggtgggg gctgggcgtg gtcatgtatg
     3241 agatgatgtg tggccgcctg cccttctaca accaggacca cgagaagctg ttcgagctga
     3301 tcctcatgga ggagatccgc ttcccgcgca cactcggccc tgaggccaag tccctgctct
     3361 ccgggctgct caagaaggac cctacacaga ggctcggtgg gggctctgag gatgccaagg
     3421 agatcatgca gcaccggttc tttgccaaca tcgtgtggca ggatgtgtat gagaagaagc
     3481 tgagcccacc tttcaagccc caggtcacct ctgagactga caccaggtat ttcgatgagg
     3541 agttcacagc tcagatgatc accatcacgc cgcctgatca agatgacagc atggagtgtg
     3601 tggacagtga gcggaggccg cacttccccc agttctccta ctcagccagt ggcacagcct
     3661 acccatacga cgtcccagac tacgcttgaa agggtgggcg cgccgaccca gctttcttgt
     3721 acaaagtggt tgatatccag cacagtggcg gccgctcgag tctagagggc ccgcggttcg
     3781 aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggttagtaat
     3841 gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3901 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3961 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4021 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     4081 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     4141 cgccctgtag cggcgcatta aataacttcg tataatgtat gctatacgaa gttatatcgt
     4201 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     4261 acacacattc cacagaattc cttgaaatac atccacaggt acacctccaa ttgactcaaa
     4321 tgatgtcaat tagtctatca gaagcttcta aagccatgac atcattttct ggaattttcc
     4381 aagctgttta aaggcacagt caacttagtg tatgtaaact tctgacccac tggaattgtg
     4441 atacagtgaa ttataagtga aataatctgt ctgtaaacaa ttgttggaaa aatgacttgt
     4501 gtcatgcaca aagtagatgt cctaactgac ttgccaaaac tattgtttgt taacaagaaa
     4561 tttgtggagt agttgaaaaa cgagttttaa tgactccaac ttaagtgtat gtaaacttcc
     4621 gacttcaact gtagggtacc gagctcgaat tcactggccg tcgttttaca acgtcgtgac
     4681 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     4741 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     4801 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4861 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4921 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4981 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5041 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5101 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5161 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5221 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5281 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5341 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5401 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5461 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5521 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5581 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5641 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5701 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5761 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5821 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5881 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5941 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6001 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6061 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6121 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6181 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6241 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6301 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6361 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6421 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6481 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6541 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6601 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6661 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6721 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6781 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6841 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6901 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6961 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7021 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     7081 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     7141 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     7201 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     7261 agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag acctatacag
     7321 ttgaagtcgg aagtttacat acacttaagt tggagtcatt aaaactcgtt tttcaactac
     7381 tccacaaatt tcttgttaac aaacaatagt tttggcaagt cagttaggac atctactttg
     7441 tgcatgacac aagtcatttt tccaacaatt gtttacagac agattatttc acttataatt
     7501 cactgtatca caattccagt gggtcagaag tttacataca ctaagttgac tgtgccttta
     7561 aacagcttgg aaaattccag aaaatgatgt catggcttta gaagcttctg atagactaat
     7621 tgacatcatt tgagtcaatt ggaggtgtac ctgtggatgt atttcaaggg gatccagaca
     7681 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagcgg ccgctagctc
     7741 gagatcttaa ttaaactagt atcgatataa cttcgtataa tgtatgctat acgaagttat
     7801 atgtacgggc cagatatacg cgttgacatt gattattgac taggcttttg caaaaagctt
     7861 tgcaaagatg gataaagttt taaacagaga ggaatctttg cagctaatgg accttctagg
     7921 tcttgaaagg agtgggaatt
//
