LOCUS       Exported                7864 bp ds-DNA     circular SYN 04-JUN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pFRTpsiCHECK_gaussia_luciferase
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7864)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 8, 2020 from SnapGene Viewer 2.7.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7864
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        73..430
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      281..416
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     intron          500..632
                     /note="chimeric intron"
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        677..695
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             701..1258
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /note="Gaussia luciferase"
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     CDS             701..1258
                     /codon_start=1
                     /product="enhanced variant of secreted Gaussia luciferase"
                     /note="Gaussia luciferase"
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     polyA_signal    1274..1322
                     /note="synthetic polyadenylation signal"
     promoter        1331..2082
                     /note="HSV TK promoter"
                     /note="herpes simplex virus thymidine kinase promoter"
     misc_feature    2118..3770
                     /note="Synthetic Firefly luciferase"
     misc_feature    2118..3770
                     /note="Synthetic Firefly luciferase"
     polyA_signal    3822..4046
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    4330..4377
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4385..5407
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /note="HygR"
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5537..5658
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5707..5723)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5731..5747
                     /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)."
     promoter        complement(5755..5785)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    5800..5821
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6109..6697)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6868..7728)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7729..7833)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
       61 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
      121 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      181 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      241 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
      301 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
      361 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
      421 cttttgcaaa aagcttgatt cttctgacac aacagtctcg aacttaagct gcagaagttg
      481 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
      541 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
      601 tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc
      661 ttaaggctag agtacttaat acgactcact ataggctagc atgggagtca aagttctgtt
      721 tgccctgatc tgcatcgctg tggccgaggc caagcccacc gagaacaacg aagacttcaa
      781 catcgtggcc gtggccagca acttcgcgac cacggatctc gatgctgacc gcgggaagtt
      841 gcccggcaag aagctgccgc tggaggtgct caaagagctg gaagccaatg cccggaaagc
      901 tggctgcacc aggggctgtc tgatctgcct gtcccacatc aagtgcacgc ccaagatgaa
      961 gaagttcatc ccaggacgct gccacaccta cgaaggcgac aaagagtccg cacagggcgg
     1021 cataggcgag gcgatcgtcg acattcctga gattcctggg ttcaaggact tggagcccct
     1081 ggagcagttc atcgcacagg tcgatctgtg tgtggactgc acaactggct gcctcaaagg
     1141 gcttgccaac gtgcagtgtt ctgacctgct caagaagtgg ctgccgcaac gctgtgcgac
     1201 ctttgccagc aagatccagg gccaggtgga caagatcaag ggggccggtg gtgactaagc
     1261 ggccgctggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     1321 tgaggatcta aatgagtctt cggacctcgc gggggccgct taagcggtgg ttagggtttg
     1381 tctgacgcgg ggggaggggg aaggaacgaa acactctcat tcggaggcgg ctcggggttt
     1441 ggtcttggtg gccacgggca cgcagaagag cgccgcgatc ctcttaagca cccccccgcc
     1501 ctccgtggag gcgggggttt ggtcggcggg tggtaactgg cgggccgctg actcgggcgg
     1561 gtcgcgcgcc ccagagtgtg accttttcgg tctgctcgca gacccccggg cggcgccgcc
     1621 gcggcggcga cgggctcgct gggtcctagg ctccatgggg accgtatacg tggacaggct
     1681 ctggagcatc cgcacgactg cggtgatatt accggagacc ttctgcggga cgagccgggt
     1741 cacgcggctg acgcggagcg tccgttgggc gacaaacacc aggacggggc acaggtacac
     1801 tatcttgtca cccggaggcg cgagggactg caggagcttc agggagtggc gcagctgctt
     1861 catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc ccggaagaaa tatatttgca
     1921 tgtctttagt tctatgatga cacaaacccc gcccagcgtc ttgtcattgg cgaattcgaa
     1981 cacgcagatg cagtcggggc ggcgcggtcc caggtccact tcgcatatta aggtgacgcg
     2041 tgtggcctcg aacaccgagc gaccctgcag cgacccgctt aaaagcttgg cattccggta
     2101 ctgttggtaa agccaccatg gccgatgcta agaacattaa gaagggccct gctcccttct
     2161 accctctgga ggatggcacc gctggcgagc agctgcacaa ggccatgaag aggtatgccc
     2221 tggtgcctgg caccattgcc ttcaccgatg cccacattga ggtggacatc acctatgccg
     2281 agtacttcga gatgtctgtg cgcctggccg aggccatgaa gaggtacggc ctgaacacca
     2341 accaccgcat cgtggtgtgc tctgagaact ctctgcagtt cttcatgcca gtgctgggcg
     2401 ccctgttcat cggagtggcc gtggcccctg ctaacgacat ttacaacgag cgcgagctgc
     2461 tgaacagcat gggcatttct cagcctaccg tggtgttcgt gtctaagaag ggcctgcaga
     2521 agatcctgaa cgtgcagaag aagctgccta tcatccagaa gatcatcatc atggactcta
     2581 agaccgacta ccagggcttc cagagcatgt acacattcgt gacatctcat ctgcctcctg
     2641 gcttcaacga gtacgacttc gtgccagagt ctttcgacag ggacaaaacc attgccctga
     2701 tcatgaacag ctctgggtct accggcctgc ctaagggcgt ggccctgcct catcgcaccg
     2761 cctgtgtgcg cttctctcac gcccgcgacc ctattttcgg caaccagatc atccccgaca
     2821 ccgctattct gagcgtggtg ccattccacc acggcttcgg catgttcacc accctgggct
     2881 acctgatttg cggctttcgg gtggtgctga tgtaccgctt cgaggaggag ctgttcctgc
     2941 gcagcctgca agactacaaa attcagtctg ccctgctggt gccaaccctg ttcagcttct
     3001 tcgctaagag caccctgatc gacaagtacg acctgtctaa cctgcacgag attgcctctg
     3061 gcggcgcccc actgtctaag gaggtgggcg aagccgtggc caagcgcttt catctgccag
     3121 gcatccgcca gggctacggc ctgaccgaga caaccagcgc cattctgatt accccagagg
     3181 gcgacgacaa gcctggcgcc gtgggcaagg tggtgccatt cttcgaggcc aaggtggtgg
     3241 acctggacac cggcaagacc ctgggagtga accagcgcgg cgagctgtgt gtgcgcggcc
     3301 ctatgattat gtccggctac gtgaataacc ctgaggccac aaacgccctg atcgacaagg
     3361 acggctggct gcactctggc gacattgcct actgggacga ggacgagcac ttcttcatcg
     3421 tggaccgcct gaagtctctg atcaagtaca agggctacca ggtggcccca gccgagctgg
     3481 agtctatcct gctgcagcac cctaacattt tcgacgccgg agtggccggc ctgcccgacg
     3541 acgatgccgg cgagctgcct gccgccgtcg tcgtgctgga acacggcaag accatgaccg
     3601 agaaggagat cgtggactat gtggccagcc aggtgacaac cgccaagaag ctgcgcggcg
     3661 gagtggtgtt cgtggacgag gtgcccaagg gcctgaccgg caagctggac gcccgcaaga
     3721 tccgcgagat cctgatcaag gctaagaaag gcggcaagat cgccgtgtaa taattctagt
     3781 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3841 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3901 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3961 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     4021 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     4081 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4141 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4201 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4261 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4321 acgtacctag aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcctt
     4381 ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     4441 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     4501 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     4561 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
     4621 tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     4681 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga
     4741 tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     4801 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     4861 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     4921 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     4981 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     5041 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     5101 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     5161 gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     5221 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     5281 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
     5341 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     5401 ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5461 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5521 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5581 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5641 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5701 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5761 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5821 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5881 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5941 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6001 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6061 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6121 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6181 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6241 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6301 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6361 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6421 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6481 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6541 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6601 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6661 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6721 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6781 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6841 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6901 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6961 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7021 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7081 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7141 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7201 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7261 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7321 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7381 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7441 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7501 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7561 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7621 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7681 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7741 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7801 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7861 cgtc
//
