LOCUS       Exported                8881 bp ds-DNA    circular SYN 09-12-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    gag-MLH1dn
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8881)
  AUTHORS   G.Yu
  TITLE     Direct Submission
  JOURNAL   Exported 2022¤¤¤Å¤¤ 12 9¤·¤Ó¤© ¤¡¤Ñ¤±¤·¤Ë¤·¤Ó¤© from SnapGene 2.3.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8881
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(174..762)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(933..1793)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1:  933 .. 1724 / #ccffcc
                        2: 1725 .. 1793 / #ccffcc / signal sequence
                     Cleavage site after base 1724"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1794..1898)
                     /gene="bla"
                     /note="AmpR promoter"
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(1924..2379)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     2521..2537
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        2547..2565
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     enhancer        2686..3065
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        3066..3269
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     intron          3381..3953
                     /note="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
                     /note="color: #------"
     CDS             4051..4467
                     /note="gag (truncated)"
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
                     /note="color: #993366; direction: RIGHT"
     CDS             5680..5706
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /note="NES"
                     /note="color: #993366"
                     /translation="LPPLERLTL"
     CDS             5719..5745
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /note="NES"
                     /note="color: #993366"
                     /translation="LPPLERLTL"
     CDS             5758..5784
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /note="NES"
                     /note="color: #993366"
                     /translation="LPPLERLTL"
     CDS             5785..5814
                     /codon_start=1
                     /note="Protease cleavate site"
                     /note="color: #ff0000
                     Cleavage site after base 5799"
                     /translation="TSTLLMENSS"
     CDS             5818..5841
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /note="FLAG"
                     /note="color: #cc99b2"
                     /translation="DYKDDDDK"
     CDS             5908..8163
                     /codon_start=1
                     /note="hMLH1dn_codon_optimized"
                     /note="color: #008080"
                     /translation="SFVAGVIRRLDETVVNRIAAGEVIQRPANAIKEMIENCLDAKSTS
                     IQVIVKEGGLKLIQIQDNGTGIRKEDLDIVCERFTTSKLQSFEDLASISTYGFRGEALA
                     SISHVAHVTITTKTADGKCAYRASYSDGKLKAPPKPCAGNQGTQITVEDLFYNIATRRK
                     ALKNPSEEYGKILEVVGRYSVHNAGISFSVKKQGETVADVRTLPNASTVDNIRSIFGNA
                     VSRELIEIGCEDKTLAFKMNGYISNANYSVKKCIFLLFINHRLVESTSLRKAIETVYAA
                     YLPKNTHPFLYLSLEISPQNVDVNVHPTKHEVHFLHEESILERVQQHIESKLLGSNSSR
                     MYFTQTLLPGLAGPSGEMVKSTTSLTSSSTSGSSDKVYAHQMVRTDSREQKLDAFLQPL
                     SKPLSSQPQAIVTEDKTDISSGRARQQDEEMLELPAPAEVAAKNQSLEGDTTKGTSEMS
                     EKRGPTSSNPRKRHREDSDVEMVEDDSRKEMTAACTPRRRIINLTSVLSLQEEINEQGH
                     EVLREMLHNHSFVGCVNPQWALAQHQTKLYLLNTTKLSEELFYQILIYDFANFGVLRLS
                     EPAPLFDLAMLALDSPESGWTEEDGPKEGLAEYIVEFLKKKAEMLADYFSLEIDEEGNL
                     IGLPLLIDNYVPPLEGLPIFILRLATEVNWDEEKECFESLSKECAMFYSIRKQYISEES
                     TLSGQQSEVPGSIPNSWKWTVEHIVYKALRSHILPPKHFTEDGNILQLANLPDLYKVF"
     polyA_signal    8240..8295
                     /note="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
                     /note="color: #a6acb3"
     primer_bind     complement(8653..8669)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    8677..8693
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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)."
                     /note="color: #31849b"
     promoter        complement(8701..8731)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 8701 .. 8707 / #ffffff / -10
                        2: 8708 .. 8725 / #ffffff
                        3: 8726 .. 8731 / #ffffff / -35"
     protein_bind    8746..8767
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
ORIGIN
        1 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
       61 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      121 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      181 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      241 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      301 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      361 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      421 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      481 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      541 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      601 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      661 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      721 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      781 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      841 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      901 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
      961 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1021 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1081 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1141 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1201 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1261 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1321 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1381 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1441 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1501 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1561 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1621 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1681 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1741 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1801 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1861 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1921 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1981 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     2041 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     2101 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     2161 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2221 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2281 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2341 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     2401 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2461 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2521 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttggagctcc
     2581 accgcggtgg cggccgctct agagagcttg gcccattgca tacgttgtat ccatatcata
     2641 atatgtacat ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga
     2701 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     2761 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     2821 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     2881 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     2941 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     3001 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     3061 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     3121 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     3181 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     3241 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
     3301 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggtcga
     3361 ccgatcctga gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta
     3421 ttgtaaaatt catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa
     3481 gatgtccctt gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct
     3541 gttgacaacc attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact
     3601 ttagcttgca tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt
     3661 actttctcta atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac
     3721 agttttagag aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt
     3781 ctggctggcg tggaaatatt cttattggta gaaacaacta caccctggtc atcatcctgc
     3841 ctttctcttt atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt
     3901 ccaaaccggg cccctctgct aaccatgttc atgccttctt ctctttccta cagctcctgg
     3961 gcaacgtgct ggttgttgtg ctgtctcatc attttggcaa agaattcctc gagggccgga
     4021 tccgaattcc tgtatttgtc tgaaaacatg ggccagactg ttaccactcc cttaagtttg
     4081 accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag
     4141 aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg
     4201 cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct
     4261 ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt
     4321 gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca
     4381 tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat
     4441 ccagccctca ctccttctct aggcgccaaa cctaaacctc aagttctttc tgacagtggg
     4501 gggccgctca tcgacctact tacagaagac cccccgcctt atagggaccc aagaccaccc
     4561 ccttccgaca gggacggaaa tggtggagaa gcgacccctg cgggagaggc accggacccc
     4621 tccccaatgg catctcgcct acgtgggaga cgggagcccc ctgtggccga ctccactacc
     4681 tcgcaggcat tccccctccg cgcaggagga aacggacagc ttcaatactg gccgttctcc
     4741 tcttctgacc tttacaactg gaaaaataat aacccttctt tttctgaaga tccaggtaaa
     4801 ctgacagctc tgatcgagtc tgtcctcatc acccatcagc ccacctggga cgactgtcag
     4861 cagctgttgg ggactctgct gaccggagaa gaaaaacaac gggtgctctt agaggctaga
     4921 aaggcggtgc ggggcgatga tgggcgcccc actcaactgc ccaatgaagt cgatgccgct
     4981 tttcccctcg agcgcccaga ctgggattac accacccagg caggtaggaa ccacctagtc
     5041 cactatcgcc agttgctcct agcgggtctc caaaacgcgg gcagaagccc caccaatttg
     5101 gccaaggtaa aaggaataac acaagggccc aatgagtctc cctcggcctt cctagagaga
     5161 cttaaggaag cctatcgcag gtacactcct tatgaccctg aggacccagg gcaagaaact
     5221 aatgtgtcta tgtctttcat ttggcagtct gccccagaca ttgggagaaa gttagagagg
     5281 ttagaagatt taaaaaacaa gacgcttgga gatttggtta gagaggcaga aaagatcttt
     5341 aataaacgag aaaccccgga agaaagagag gaacgtatca ggagagaaac agaggaaaaa
     5401 gaagaacgcc gtaggacaga ggatgagcag aaagagaaag aaagagatcg taggagacat
     5461 agagagatga gcaagctatt ggccactgtc gttagtggac agaaacagga tagacaggga
     5521 ggagaacgaa ggaggtccca actcgatcgc gaccagtgtg cctactgcaa agaaaagggg
     5581 cactgggcta aagattgtcc caagaaacca cgaggacctc ggggaccaag accccagacc
     5641 tccctcctga ccctagatga ctctggcggc tcacttcaac tgcctccact tgaaagactg
     5701 acactgggat cattacaatt acctccttta gaacgattaa cactcggttc actacagctt
     5761 ccgcctcttg agagattgac attaacgtcc acgctgctaa tggagaactc gtctggagat
     5821 tacaaagacg atgacgataa gaaacgtacg gccgacggaa gcgagttcga gtcaccaaag
     5881 aagaagcgga aagtcagcgg ggggtcaagc ttcgttgctg gagtcatccg gagactggac
     5941 gagacagtgg tgaacagaat tgccgccggc gaggtgatcc agagacctgc caatgcaatc
     6001 aaggagatga tcgagaactg tctggacgcc aagtccacaa gcattcaggt gatcgtgaag
     6061 gagggcggac tgaagctgat ccagatccaa gacaacggca caggcatcag aaaggaagat
     6121 ctggacatcg tgtgtgaacg gttcaccaca tctaagctgc agtcttttga ggatctggcc
     6181 tctatcagta cctacggctt cagaggcgag gccctggcca gcatcagcca cgtggcccat
     6241 gtgaccatca ccaccaaaac cgccgacggc aaatgcgctt atcgcgctag ctacagcgac
     6301 ggcaagctga aagccccgcc aaagccttgc gccggcaacc agggtacaca gataacagtg
     6361 gaggatctgt tctacaacat cgccacccgg agaaaggccc tgaaaaatcc cagcgaggag
     6421 tacggcaaga tcctggaagt cgtcggcaga tactccgtgc acaacgccgg aatcagcttt
     6481 agcgtaaaga agcagggaga aaccgtggcc gatgtgcgca ccctgccaaa tgccagcacc
     6541 gtggataaca tcagaagcat tttcggaaat gccgtgtcca gagaactgat cgagatcggc
     6601 tgcgaagata agaccctggc ttttaagatg aacggctaca tcagcaacgc caattactct
     6661 gtgaagaagt gcatctttct tctgttcatc aaccacagac tggtggaaag caccagcctg
     6721 cggaaagcca tcgagacagt gtacgccgcc tacctgccta agaacaccca ccccttcctg
     6781 tacctgagcc tcgagatcag ccctcagaac gtggacgtca atgtgcatcc tacaaagcac
     6841 gaggtgcact tcctgcacga ggaaagcatc ctggaaagag tgcagcagca cattgagagc
     6901 aagctgctgg gctctaacag cagcagaatg tacttcacac agaccctgct gcctggcctg
     6961 gccggcccct caggcgaaat ggttaagtcc acaacctctc tgacctcatc cagcaccagc
     7021 ggttcttccg ataaggtgta cgcccaccag atggtgcgga ccgactctcg ggagcagaag
     7081 ctggacgcct ttctgcaacc tctgagcaaa cctctgagct ctcagcctca ggccatcgtg
     7141 accgaggaca agacagatat ctcctccggc cgtgccagac agcaggacga agaaatgctc
     7201 gagctgccag ctcctgccga ggtggccgcc aagaaccaga gcctggaggg agataccaca
     7261 aagggcacca gcgaaatgag cgagaagcgg ggccctacct ccagcaaccc cagaaaacgg
     7321 caccgggagg acagcgacgt ggaaatggtg gaggacgaca gcagaaagga aatgacagcc
     7381 gcttgtaccc ctagaagaag aatcatcaac ctgacctccg tgctgagcct gcaggaggag
     7441 atcaacgagc agggccacga ggtgctgaga gagatgctgc acaatcacag cttcgtgggc
     7501 tgcgtgaacc ctcaatgggc cctggctcag catcaaacaa agctgtacct gctgaacacc
     7561 accaagctga gcgaagagct gttctaccag atcctcatct acgacttcgc caacttcggc
     7621 gtgctacgcc tgagcgagcc cgcccctctg tttgacctgg ccatgctggc tctggatagc
     7681 ccagaaagcg gctggacaga agaggacgga cctaaagagg ggctggctga atacatcgtg
     7741 gagttcctga agaaaaaggc cgagatgctg gccgactact tttctctgga aatcgacgag
     7801 gaaggcaacc tgatcggcct gcctctgctg atcgataact acgtgcctcc cctggaaggc
     7861 ctgcccatct tcatcctgag actggctaca gaggtgaact gggacgagga aaaggaatgc
     7921 ttcgagtctc tgagcaagga gtgcgccatg ttctatagca tcagaaaaca gtacatctct
     7981 gaagagagca ctctgtctgg ccagcagagt gaagtgcccg gaagcatccc caacagctgg
     8041 aagtggaccg tggaacacat cgtgtacaag gccctgcgga gccacattct ccctcctaag
     8101 cacttcaccg aggacggcaa catcctgcag ctggccaacc tgcccgacct ttataaggtt
     8161 ttctaaaccg gtgatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     8221 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     8281 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     8341 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     8401 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     8461 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     8521 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     8581 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacgg atcggccgca
     8641 attcgtaacc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     8701 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     8761 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     8821 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     8881 t
//
