LOCUS       Exported                7330 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 7330)
  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..7330
                     /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"
     primer_bind     769..789
                     /label=YA0182
     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             1025..1741
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1213..1232)
                     /label=GFP Reverse
     misc_feature    1757..3154
                     /label=G3BP1
     CDS             2201..2203
                     /codon_start=1
                     /label=S149A (tct to gct)
                     /translation="S"
     misc_feature    2747..3154
                     /label=Region deleted for Delta RBP
     polyA_signal    3288..3512
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3796..3843
                     /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             3851..4873
                     /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    5003..5124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5173..5189)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5197..5213
                     /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(5221..5251)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5266..5287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5575..6163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6334..7194)
                     /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(7195..7299)
                     /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 cttttagtga accgtcagat ccgctagcgc taccggtcgc
     1021 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1081 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1141 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1201 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1261 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1321 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1381 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1441 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1501 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1561 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1621 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1681 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1741 gtccggactc agatctatgg tgatggagaa gcctagtccc ctgctggtcg ggcgggaatt
     1801 tgtgagacag tattacacac tgctgaacca ggccccagac atgctgcata gattttatgg
     1861 aaagaactct tcttatgtcc atgggggatt ggattcaaat ggaaagccag cagatgcagt
     1921 ctacggacag aaagaaatcc acaggaaagt gatgtcacaa aacttcacca actgccacac
     1981 caagattcgc catgttgatg ctcatgccac gctaaatgat ggtgtggtag tccaggtgat
     2041 ggggcttctc tctaacaaca accaggcttt gaggagattc atgcaaacgt ttgtccttgc
     2101 tcctgagggg tctgttgcaa ataaattcta tgttcacaat gatatcttca gataccaaga
     2161 tgaggtcttt ggtgggtttg tcactgagcc tcaggaggag tctgaagaag aagtagagga
     2221 acctgaagaa agacagcaaa cacctgaggt ggtacctgat gattctggaa ctttctatga
     2281 tcaggcagtt gtcagtaatg acatggaaga acatttagag gagcctgttg ctgaaccaga
     2341 gcctgatcct gaaccagaac cagaacaaga acctgtatct gaaatccaag aggaaaagcc
     2401 tgagccagta ttagaagaaa ctgcccctga ggatgctcag aagagttctt ctccagcacc
     2461 tgcagacata gctcagacag tacaggaaga cttgaggaca ttttcttggg catctgtgac
     2521 cagtaagaat cttccaccca gtggagctgt tccagttact gggataccac ctcatgttgt
     2581 taaagtacca gcttcacagc cccgtccaga gtctaagcct gaatctcaga ttccaccaca
     2641 aagacctcag cgggatcaaa gagtgcgaga acaacgaata aatattcctc cccaaagggg
     2701 acccagacca atccgtgagg ctggtgagca aggtgacatt gaaccccgaa gaatggtgag
     2761 acaccctgac agtcaccaac tcttcattgg caacctgcct catgaagtgg acaaatcaga
     2821 gcttaaagat ttctttcaaa gttatggaaa cgtggtggag ttgcgcatta acagtggtgg
     2881 gaaattaccc aattttggtt ttgttgtgtt tgatgattct gagcctgttc agaaagtcct
     2941 tagcaacagg cccatcatgt tcagaggtga ggtccgtctg aatgtcgaag agaagaagac
     3001 tcgagctgcc agggaaggcg accgacgaga taatcgcctt cggggacctg gaggccctcg
     3061 aggtgggctg ggtggtggaa tgagaggccc tccccgtgga ggcatggtgc agaaaccagg
     3121 atttggagtg ggaagggggc ttgcgccacg gcagtgagaa ttctgcagtc gacggtaccg
     3181 cgggcccggg atccactagt ccagtgtggt ggaattctgc agatatccag cacagtggcg
     3241 gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3301 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3361 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3421 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3481 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3541 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3601 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3661 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3721 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3781 tggttcacgt acctagaagt tcctattccg aagttcctat tctctagaaa gtataggaac
     3841 ttccttggcc aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     3901 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     3961 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     4021 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     4081 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     4141 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     4201 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     4261 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     4321 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     4381 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     4441 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     4501 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     4561 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     4621 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     4681 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     4741 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     4801 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     4861 ggcaaaggaa tagcacgtac tacgagattt cgattccacc gccgccttct atgaaaggtt
     4921 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4981 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5041 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5101 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5161 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5221 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5281 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5341 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5401 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5461 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5521 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5581 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5641 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5701 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5761 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5821 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5881 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5941 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6001 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6061 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6121 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6181 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6241 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6301 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6361 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6421 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6481 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6541 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6601 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6661 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6721 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6781 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6841 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6901 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6961 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7021 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7081 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7141 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7201 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7261 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7321 acctgacgtc
//
