LOCUS       Exported                5746 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    HA-GbL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5746)
  AUTHORS   Sarbassov DD, Ali SM, Kim DH, Guertin DA, Latek RR, 
            Erdjument-Bromage H, Tempst P, Sabatini DM
  TITLE     Rictor, a novel binding partner of mTOR, defines a 
            rapamycin-insensitive and raptor-independent pathway that regulates 
            the cytoskeleton.
  JOURNAL   Curr Biol 2004 Jul 27;14(14):1296-302.
  PUBMED    15268862
REFERENCE   2  (bases 1 to 5746)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5746
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      25..613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     514..533
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     767..784
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    901..922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        937..967
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    975..991
                     /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)."
     primer_bind     980..1002
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     999..1015
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     999..1015
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        1044..1423
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1424..1627
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1578..1598
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1624..1648
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1858..1876
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1858..1875
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1889..1908
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1993..2002
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2005..2031
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3076..3085
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    3089..3223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3126..3145)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3180..3199
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3292..3649
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3315..3335)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3500..3635
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3562..3581
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3669..3686)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3669..3685)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3678..3700)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3898..4353
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3985..4004)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4195..4216
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4370..4389)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4489..4511
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4549..4567)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4635..4739
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4740..5600
                     /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     complement(4958..4977)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
       61 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      121 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
      181 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      241 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      301 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      361 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      421 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
      481 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      541 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      601 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      661 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      721 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
      781 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
      841 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
      901 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
      961 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
     1021 tacgaattaa ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat
     1081 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     1141 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     1201 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     1261 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     1321 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     1381 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     1441 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     1501 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     1561 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     1621 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
     1681 ccatagaaga caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg
     1741 gattccccgt gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt
     1801 ggcttcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     1861 taggtgacac tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag
     1921 gtccaactgc acctcggttc tatcgattga attcccgaac cgacagtcgg tctcttcacc
     1981 aaggccattc gcgccaccat gggctacccc tatgatgtgc ctgactacgc agatctcaat
     2041 ggtggtggtg gtgggtcgac gatgaacacc tccccaggca cggtgggcag tgacccggtc
     2101 atcctggcca ctgcaggcta cgaccacacc gtgcgcttct ggcaggccca cagcggcatc
     2161 tgcacccgga cggtgcagca ccaggactcc caggtgaatg ccttggaggt cacaccggac
     2221 cgcagcatga ttgctgctgc aggttaccag cacatccgca tgtatgatct caactccaat
     2281 aaccctaacc ccatcatcag ctacgacggc gtcaacaaga acatcgcgtc tgtgggcttc
     2341 cacgaagacg gccgctggat gtacacgggc ggcgaggact gcacagccag gatctgggac
     2401 ctcaggtccc ggaacctgca gtgccagcgg atcttccagg tgaacgcacc cattaactgc
     2461 gtgtgcctgc accccaacca ggcagagctc atcgtgggtg accagagcgg ggctatccac
     2521 atctgggact tgaaaacaga ccacaacgag cagctgatcc ctgagcccga ggtctccatc
     2581 acgtccgccc acatcgatcc cgacgccagc tacatggcag ctgtcaatag caccggaaac
     2641 tgctatgtct ggaatctgac ggggggcatt ggtgacgagg tgacccagct catccccaag
     2701 actaagatcc ctgcccacac gcgctacgcc ctgcagtgtc gcttcagccc cgactccacg
     2761 ctcctcgcca cctgctcggc tgatcagacg tgcaagatct ggaggacgtc caacttctcc
     2821 ctgatgacgg agctgagcat caagagcggc aaccccgggg agtcctcccg cggctggatg
     2881 tggggctgcg ccttctcggg ggactcccag tacatcgtca ctgcttcctc ggacaacctg
     2941 gcccggctct ggtgtgtgga gactggagag atcaagagag agtatggcgg ccaccagaag
     3001 gctgttgtct gcctggcctt caatgacagt gtgctgggct aggcggccgc taagtaagta
     3061 aggatcccca gcttggccgc catggcccaa cttgtttatt gcagcttata atggttacaa
     3121 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3181 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcgggaatt aattcggcgc
     3241 agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg
     3301 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3361 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3421 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3481 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3541 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3601 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctgttaac
     3661 agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     3721 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     3781 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     3841 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3901 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3961 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4021 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4081 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4141 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4201 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     4261 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4321 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4381 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4441 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4501 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4561 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4621 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4681 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4741 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4801 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4861 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4921 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4981 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5041 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5101 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5161 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5221 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5281 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5341 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5401 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5461 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5521 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5581 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5641 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5701 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccc
//
