LOCUS       Exported                9447 bp ds-DNA     circular SYN 05-NOV-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9447)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Nov 5, 2019 from SnapGene Viewer 4.3.10
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9447
                     /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"
     intron          1016..1148
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1193..1211
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1227..1236
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1233..1949
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1963..5319
                     /label=UPF1
     misc_feature    3772..3774
                     /label=R615A
     misc_feature    3868..3873
                     /label=DE647AA Helicase/ATPase Mutant
     polyA_signal    5405..5629
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    5913..5960
                     /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             5968..6990
                     /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    7120..7241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7290..7306)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7314..7330
                     /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(7338..7368)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7383..7404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7692..8280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8451..9311)
                     /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(9312..9416)
                     /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 cttcttgcag aagttggtcg tgaggcactg ggcaggtaag
     1021 tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag
     1081 aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct
     1141 ctccacaggt gtccactccc agttcaatta cagctcttaa ggctagagta cttaatacga
     1201 ctcactatag gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca
     1261 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1321 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1381 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1441 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1501 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1561 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1621 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1681 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1741 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1801 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1861 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1921 tcactctcgg catggacgag ctgtacaagc ttagcgtgga agcttagcgt ggaggcgtac
     1981 gggcccagct cgcagactct cactttcctg gacacggagg aggccgagct gcttggcgcc
     2041 gacacacagg gctccgagtt cgagttcacc gactttactc ttcctagcca gacgcagacg
     2101 ccccccggcg gccccggcgg cccgggcggt ggcggcgcgg gaggcccggg cggcgcgggc
     2161 gcgggcgctg cggcgggaca gctcgacgcg caggttgggc ccgaaggcat cctgcagaac
     2221 ggggctgtgg acgacagtgt agccaagacc agccagttgt tggctgagtt gaacttcgag
     2281 gaagatgaag aagacaccta ttacacgaag gacctcccca tacacgcctg cagttactgt
     2341 ggaatacacg atcctgcctg cgtggtttac tgtaatacca gcaagaagtg gttctgcaac
     2401 ggacgtggaa atacttctgg cagccacatt gtaaatcacc ttgtgagggc aaaatgcaaa
     2461 gaggtgaccc tgcacaagga cgggcccctg ggggagacag tcctggagtg ctacaactgc
     2521 ggctgtcgca acgtcttcct cctcggcttc atcccggcca aagctgactc agtggtggtg
     2581 ctgctgtgca ggcagccctg tgccagccag agcagcctca aggacatcaa ctgggacagc
     2641 tcgcagtggc agccgctgat ccaggaccgc tgcttcctgt cctggctggt caagatcccc
     2701 tccgagcagg agcagctgcg ggcacgccag atcacggcac agcagatcaa caagctggag
     2761 gagctgtgga aggaaaaccc ttctgccacg ctggaggacc tggagaagcc gggggtggac
     2821 gaggagccgc agcatgtcct cctgcggtac gaggacgcct accagtacca gaacatattc
     2881 gggcccctgg tcaagctgga ggccgactac gacaagaagc tgaaggagtc ccagactcaa
     2941 gataacatca ctgtcaggtg ggacctgggc cttaacaaga agagaatcgc ctacttcact
     3001 ttgcccaaga ctgactctga catgcggctc atgcaggggg atgagatatg cctgcggtac
     3061 aaaggggacc ttgcgcccct gtggaaaggg atcggccacg tcatcaaggt ccctgataat
     3121 tatggcgatg agatcgccat tgagctgcgg agcagcgtgg gtgcacctgt ggaggtgact
     3181 cacaacttcc aggtggattt tgtgtggaag tcgacctcct ttgacaggat gcagagcgca
     3241 ttgaaaacgt ttgccgtgga tgagacctcg gtgtctggct acatctacca caagctgttg
     3301 ggccacgagg tggaggacgt aatcatcaag tgccagctgc ccaagcgctt cacggcgcag
     3361 ggcctccccg acctcaacca ctcccaggtt tatgccgtga agactgtgct gcaaagacca
     3421 ctgagcctga tccagggccc gccaggcacg gggaagacgg tgacgtcggc caccatcgtc
     3481 taccacctgg cccggcaagg caacgggccg gtgctggtgt gtgctccgag caacatcgcc
     3541 gtggaccagc taacggagaa gatccaccag acggggctaa aggtcgtgcg cctctgcgcc
     3601 aagagccgtg aggccatcga ctccccggtg tcttttctgg ccctgcacaa ccagatcagg
     3661 aacatggaca gcatgcctga gctgcagaag ctgcagcagc tgaaagacga gactggggag
     3721 ctgtcgtctg ccgacgagaa gcggtaccgg gccttgaagc gcaccgcaga gagagagctg
     3781 ctgatgaacg cagatgtcat ctgctgcaca tgtgtgggcg ccggtgaccc gaggctggcc
     3841 aagatgcagt tccgctccat tttaatcgac gaaagcaccc aggccaccga gccggagtgc
     3901 atggttcccg tggtcctcgg ggccaagcag ctgatccttg taggcgacca ctgccagctg
     3961 ggcccagtgg tgatgtgcaa gaaggcggcc aaggccgggc tgtcacagtc gctcttcgag
     4021 cgcctggtgg tgctgggcat ccggcccatc cgcctgcagg tccagtaccg gatgcaccct
     4081 gcactcagcg ccttcccatc caacatcttc tacgagggct ccctccagaa tggtgtcact
     4141 gcagcggatc gtgtgaagaa gggatttgac ttccagtggc cccaacccga taaaccgatg
     4201 ttcttctacg tgacccaggg ccaagaggag attgccagct cgggcacctc ctacctgaac
     4261 aggaccgagg ctgcgaacgt ggagaagatc accacgaagt tgctgaaggc aggcgccaag
     4321 ccggaccaga ttggcatcat cacgccctac gagggccagc gctcctacct ggtgcagtac
     4381 atgcagttca gcggctccct gcacaccaag ctctaccagg aggtggagat cgccagtgtg
     4441 gacgcctttc agggacgcga gaaggacttc atcatcctgt cctgtgtgcg ggccaacgag
     4501 caccaaggca ttggcttttt aaatgacccc aggcgtctga acgtggccct gaccagagca
     4561 aggtatggcg tcatcattgt gggcaacccg aaggcactat caaagcagcc gctctggaac
     4621 cacctgctga actactataa ggagcagaag gtgctggtgg aggggccgct caacaacctg
     4681 cgtgagagcc tcatgcagtt cagcaagcca cggaagctgg tcaacactat caacccggga
     4741 gcccgcttca tgaccacagc catgtatgat gcccgggagg ccatcatccc aggctccgtc
     4801 tatgatcgga gcagccaggg ccggccttcc agcatgtact tccagaccca tgaccagatt
     4861 ggcatgatca gtgccggccc tagccacgtg gctgccatga acattcccat ccccttcaac
     4921 ctggtcatgc cacccatgcc accgcctggc tattttggac aagccaacgg gcctgctgca
     4981 gggcgaggca ccccgaaagg caagactggt cgtgggggac gccagaagaa ccgctttggg
     5041 cttcctggac ccagccagac taacctcccc aacagccaag ccagccagga tgtggcgtca
     5101 cagcccttct ctcagggcgc cctgacgcag ggctacatct ccatgagcca gccttcccag
     5161 atgagccagc ccggcctctc ccagccggag ctgtcccagg acagttacct tggtgacgag
     5221 tttaaatcac aaatcgacgt ggcgctctca caggactcca cgtaccaggg agagcgggct
     5281 taccagcatg gcggggtgac ggggctgtcc cagtattaaa aggtggcggc ggaagagcta
     5341 aagagtcgac ccgggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc
     5401 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     5461 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     5521 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     5581 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     5641 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     5701 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5761 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5821 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5881 aaacttgatt agggtgatgg ttcacgtacc tagaagttcc tattccgaag ttcctattct
     5941 ctagaaagta taggaacttc cttggccaaa aagcctgaac tcaccgcgac gtctgtcgag
     6001 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     6061 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     6121 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     6181 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc
     6241 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     6301 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     6361 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     6421 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     6481 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     6541 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     6601 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     6661 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     6721 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     6781 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     6841 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     6901 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     6961 cccagcactc gtccgagggc aaaggaatag cacgtactac gagatttcga ttccaccgcc
     7021 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     7081 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     7141 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     7201 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     7261 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     7321 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     7381 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     7441 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7501 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7561 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7621 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7681 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7741 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7801 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7861 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7921 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7981 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8041 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     8101 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     8161 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8221 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     8281 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8341 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     8401 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8461 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8521 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8581 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8641 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8701 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8761 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8821 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8881 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8941 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     9001 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     9061 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     9121 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     9181 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     9241 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     9301 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     9361 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9421 tttccccgaa aagtgccacc tgacgtc
//
