LOCUS       Exported                7018 bp DNA     circular SYN 22-JUL-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7018)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Jul 22, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7018
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             996..1019
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1020..2930
                     /codon_start=1
                     /product="I110L, D111E, D117E, A121G, G139A, Y140F, T147S,
                     R166K"
                     /label=H.S. PABPC1 (siRNA resistant v2, 8 mutations)
                     /translation="MNPSAPSYPMASLYVGDLHPDVTEAMLYEKFSPAGPILSIRVCRD
                     MITRRSLGYAYVNFQQPADAERALDTMNFDVIKGKPVRIMWSQRDPSLRKSGVGNIFIK
                     NLDKSLENKALYETFSGFGNILSCKVVCDENGSKAFGFVHFESQEAAERAIEKMNGMLL
                     NDKKVFVGRFKSRKEREAELGARAKEFTNVYIKNFGEDMDDERLKDLFGKFGPALSVKV
                     MTDESGKSKGFGFVSFERHEDAQKAVDEMNGKELNGKQIYVGRAQKKVERQTELKRKFE
                     QMKQDRITRYQGVNLYVKNLDDGIDDERLRKEFSPFGTITSAKVMMEGGRSKGFGFVCF
                     SSPEEATKAVTEMNGRIVATKPLYVALAQRKEERQAHLTNQYMQRMASVRAVPNPVINP
                     YQPAPPSGYFMAAIPQTQNRAAYYPPSQIAQLRPSPRWTAQGARPHPFQNMPGAIRPAA
                     PRPPFSTMRPASSQVPRVMSTQRVANTSTQTMGPRPAAAAAAATPAVRTVPQYKYAAGV
                     RNPQQHLNAQPQVTMQQPAVHVQGQEPLTASMLASAPPQEQKQMLGERLFPLIQAMHPT
                     LAGKITGMLLEIDNSELLHMLESPESLRSKVDEAVAVLQAHQAKEAAQKAVNSATGVPT
                     V"
     misc_feature    2931..2950
                     /label=3' end preferred cloning region
     polyA_signal    2976..3200
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3484..3531
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3539..4561
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4691..4812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4861..4877)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4885..4901
                     /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)."
     promoter        complement(4909..4939)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4954..4975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5263..5851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6022..6882)
                     /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"
     promoter        complement(6883..6987)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttgccgcca ccatggatta caaggatgac gacgataaga
     1021 tgaaccccag tgcccccagc taccccatgg cctcgctcta cgtgggggac ctccaccccg
     1081 acgtgaccga ggcgatgctc tacgagaagt tcagcccggc cgggcccatc ctctccatcc
     1141 gggtctgcag ggacatgatc acccgccgct ccttgggcta cgcgtatgtg aacttccagc
     1201 agccggcgga cgcggagcgt gctttggaca ccatgaattt tgatgttata aagggcaagc
     1261 cagtacgcat catgtggtct cagcgtgatc catcacttcg caaaagtgga gtaggcaaca
     1321 tattcattaa aaatctggat aagtctctag aaaacaaagc actgtatgaa acattttctg
     1381 gttttggtaa catcctttca tgtaaggtgg tatgcgatga aaatggttcc aaggcctttg
     1441 gatttgtaca ctttgagagc caggaagcag ctgaaagagc tattgaaaaa atgaatggaa
     1501 tgctcctaaa tgataaaaaa gtatttgttg gacgatttaa gtctcgtaaa gaacgagaag
     1561 ctgaacttgg agctagggca aaagaattca ccaatgttta catcaagaat tttggagaag
     1621 acatggatga tgagcgcctt aaggatctct ttggcaagtt tgggcctgcc ttaagtgtga
     1681 aagtaatgac tgatgaaagt ggaaaatcca aaggatttgg atttgtaagc tttgaaaggc
     1741 atgaagatgc acagaaagct gtggacgaaa tgaatggtaa ggagctcaat ggaaaacaaa
     1801 tttatgttgg tcgagctcag aaaaaggtgg aacggcagac ggaacttaag cgcaaatttg
     1861 aacagatgaa acaagatagg atcaccagat accagggtgt taatctttat gtgaaaaatc
     1921 ttgatgatgg tattgatgat gaacgtctcc ggaaagagtt ttctccattt ggtacaatca
     1981 ctagtgcaaa ggttatgatg gagggtggtc gcagcaaagg gtttggtttt gtatgtttct
     2041 cctccccaga agaagccact aaagcagtta cagaaatgaa cggtagaatt gtggccacaa
     2101 agccattgta tgtagcttta gctcagcgca aagaagagcg ccaggctcac ctcactaacc
     2161 agtatatgca gagaatggca agtgtacgag ctgttcccaa ccctgtaatc aacccctacc
     2221 agccagcacc tccttcaggt tacttcatgg cagctatccc acagactcag aaccgtgctg
     2281 catactatcc tcctagccaa attgctcaac taagaccaag tcctcgctgg actgctcagg
     2341 gtgccagacc tcatccattc caaaatatgc ccggtgctat ccgcccagct gctcctagac
     2401 caccatttag tactatgaga ccagcttctt cacaggttcc acgagtcatg tcaacacagc
     2461 gtgttgctaa cacatcaaca cagacaatgg gtccacgtcc tgcagctgca gccgctgcag
     2521 ctactcctgc tgtccgcacc gttccacagt ataaatatgc tgcaggagtt cgcaatcctc
     2581 agcaacatct taatgcacag ccacaagtta caatgcaaca gcctgctgtt catgtacaag
     2641 gtcaggaacc tttgactgct tccatgttgg catctgcccc tcctcaagag caaaagcaaa
     2701 tgttgggtga acggctgttt cctcttattc aagccatgca ccctactctt gctggtaaaa
     2761 tcactggcat gttgttggag attgataatt cagaacttct tcatatgctc gagtctccag
     2821 agtcactccg ttctaaggtt gatgaagctg tagctgtact acaagcccac caagctaaag
     2881 aggctgccca gaaagcagtt aacagtgcca ccggtgttcc aactgtttaa cgctcgagtc
     2941 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3001 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3061 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3121 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3181 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3241 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3301 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3361 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3421 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac
     3481 ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa
     3541 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     3601 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     3661 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     3721 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     3781 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     3841 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     3901 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     3961 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     4021 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     4081 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     4141 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     4201 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     4261 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     4321 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     4381 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     4441 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     4501 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     4561 gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4621 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4681 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4741 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4801 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4861 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4921 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4981 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5041 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5101 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5161 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5221 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5281 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5341 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5401 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5461 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5521 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5581 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5641 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5701 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5761 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5821 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5881 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5941 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6001 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6061 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6121 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6181 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6241 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6301 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6361 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6421 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6481 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6541 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6601 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6661 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6721 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6781 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6841 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6901 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6961 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
