LOCUS       Exported                7044 bp DNA     circular SYN 27-JAN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7044)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 28, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    237..285
                     /note="synthetic polyadenylation signal"
     CDS             complement(286..1947)
                     /codon_start=1
                     /product="secreted Cypridina luciferase"
                     /label=CLuc
                     /note="contains 30 codon substitutions for efficient 
                     translation in mammalian cells"
                     /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECIDS
                     SCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTYVL
                     GQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADPIIA
                     NPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLCGDLK
                     MMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPINFYYY
                     TISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQFQGPCK
                     EILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVGGEAVSVP
                     YSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDFS
                     DDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHKPDRVERCMY
                     EYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK"
     primer_bind     286..320
                     /label=pDY1102_2R
     primer_bind     286..318
                     /label=551_Cluc_1R
     primer_bind     286..316
                     /label=PD2469
     primer_bind     complement(1926..1954)
                     /label=551_Cluc_1F
     primer_bind     complement(1927..1954)
                     /label=pDY1102_2F
     misc_feature    complement(1948..1953)
                     /label=Kozak
     primer_bind     complement(1962..1996)
                     /label=PD5805
     promoter        complement(1973..1991)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2036..2239)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        2240..2619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2733..3910
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     2785..2802
                     /label=pDY551_MunI
     intron          2964..3901
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3900..3922
                     /label=PD5592
     primer_bind     3912..3936
                     /label=PD5804 mRNA Forward
     primer_bind     3915..3937
                     /label=NGS F
     misc_feature    3929..3934
                     /label=Kozak
     CDS             3935..3958
                     /codon_start=1
                     /label=cloning site
                     /translation="MRDHVKVS"
     CDS             3959..4024
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="GSGATNFSLLKQAGDVEENPGP"
     primer_bind     3959..3983
                     /label=PD6581
     primer_bind     complement(4003..4024)
                     /label=PD5900
     CDS             4025..4579
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="GVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRGK
                     LPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESAQ
                     GGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQR
                     CATFASKIQGQVDKIKGAGGD"
     primer_bind     4071..4091
                     /label=pd6330
     primer_bind     complement(4436..4456)
                     /label=551_IRES_cloning_R
     primer_bind     complement(4562..4592)
                     /label=PD5774 reverse mRNA
     misc_feature    4580..4628
                     /label=Poly A Signal
     primer_bind     complement(4639..4665)
                     /label=Dual Stop Reverse
     primer_bind     complement(4642..4664)
                     /label=PD6567
     primer_bind     complement(4645..4671)
                     /label=PD5318
     misc_feature    4650..4679
                     /label=EcoRI GH
     misc_feature    complement(4809..4908)
                     /label=pBR322
     misc_feature    complement(4909..5045)
                     /label=pBR322
     misc_feature    complement(5046..7000)
                     /label=pBR322
     misc_feature    5141..5245
                     /label=AmpR promoter
     promoter        5141..5245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5246..6106
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSGGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    complement(join(7001..7044,1..3))
                     /label=pBR322
ORIGIN
        1 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
       61 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
      121 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      181 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccacaca
      241 caaaaaacca acacacagat gtaatgaaaa taaagatatt ttattttatt tgcattcatc
      301 tggtacttct agggtgtctc catgctttat gtagcattct ttcttgaact cccatgcgtg
      361 gtcacagaaa ccctgttgtc ccctcaggca atactcgtac atgcatcgtt cgacacggtc
      421 aggcttgtgg cacacgttac atttctgatc aagatcactt tgaccggcga agagactatt
      481 gcagagtcgt tcagcttcag gtttctgttc ttcggtgcat cccggtgggt tgggggtcag
      541 atcacaggct ccttcagcat caaaagaatc atcactgaaa tcctggttgt agttaccgca
      601 aataccgcaa gtcttaccat cgaatggatc tctaatatgt acgacgagca gttgcttgaa
      661 gttgaacttg accaccagag cttcaggtag gatggctgta gtcagtatgt caccatcttg
      721 ccagtagatg gaagtgttct gagagctgta cgggacggac acggcttctc ctccaaccag
      781 aatctgtttt ccatcaacaa tgagttctac tacagtcgat tgtttcctga ttcgtacttt
      841 ctctacttca gtgtaagagt caacattcct gtgtgaaacc ttcacatccc aagtgttcca
      901 gaaacagtcg gcggccataa gaatctcctt gcagggaccc tggaattggt atcttgcttt
      961 gtcaaatgta tcgtagaaag tatgtccaga caaaacgcag gttcctctag tttcgggagc
     1021 agcgcacgtc tccctgtagt caagcagcac gtgtgaggct cgctcgtctc cacccataca
     1081 gcgggcgaag gcgcaggaga tggtgtagta gtagaagttg atggggttgc ggcagctgtc
     1141 cttgtacggc tccagaagac ctttgcagta tgcaacgtca tcaggatttc catagagtgg
     1201 acaaccgtca aactcctggt tgatcttagg ctgaatagcg agttgttctg gatctgaagt
     1261 gaagtctgta tcttccatca ttttaagatc tccacagagg ccagagatca ttcctttgtt
     1321 tgctgtgtct ggggcgattc ttacagatct tcctccgagg atgtcgatca cgatcagttt
     1381 gaagaactca atgacggtga tgttgaagcc tggcatctca acaacagcga tggtgacctc
     1441 gccgatggtg tacgggttgg cgatgatagg gtcagctcca ccgtttacag tgatgggatt
     1501 ctcagtagct tgagcgatgt caatgatgtc tccagccact tccagtcttg tcttggtcag
     1561 cacagccccc ttggttccat ccaggttctc cagggtgatg gacaccttcc agtcgccgcc
     1621 cttggttcct tgacccaaca cgtatgtacc aggttcctgg aactggaatc tctttccata
     1681 gaatgttcta aaccatcctg cggcctctac tctgcattca attacatact gacacattcg
     1741 gcaacatgtt tttcctggtt tattttcaca cagtccatct gatagtatgt ctctcgtgca
     1801 ggtgccacag ctgctatcaa tacattctcc ttctttagct tcacaggaag ttggaactgt
     1861 gtttggtgga tcaggttcgt aaggacagtc ctggcaatga acagtggcgc agtagactaa
     1921 tgcaacggca agaattaagg tcttcatggt ggcggtacca agcttgcgtc tccctatagt
     1981 gagtcgtatt aatttcgata agccagtaag cagtgggttc tctagttagc cagagagctc
     2041 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     2101 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     2161 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     2221 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     2281 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     2341 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     2401 tttaccgtaa atagtccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     2461 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     2521 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     2581 taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat ctggcccgta
     2641 catcgaagct gatgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat
     2701 ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc
     2761 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
     2821 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     2881 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     2941 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct
     3001 ttacgggtta tggcccttgc gtgccttgaa ttacttccac tggctgcagt acgtgattct
     3061 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
     3121 cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat
     3181 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
     3241 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
     3301 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
     3361 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
     3421 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc
     3481 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
     3541 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
     3601 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
     3661 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
     3721 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac
     3781 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
     3841 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca
     3901 ggtgtcgtga cgtacgggat cctctagagc caccatgcga gaccatgtca aggtctcagg
     3961 aagcggagct actaacttca gcctgctgaa gcaggctggc gacgtggagg agaaccctgg
     4021 acctggagtc aaagttctgt ttgccctgat ctgcatcgct gtggccgagg ccaagcccac
     4081 cgagaacaac gaagacttca acatcgtggc cgtggccagc aacttcgcga ccacggatct
     4141 cgatgctgac cgcgggaagt tgcccggcaa gaagctgccg ctggaggtgc tcaaagagat
     4201 ggaagccaat gcccggaaag ctggctgcac caggggctgt ctgatctgcc tgtcccacat
     4261 caagtgcacg cccaagatga agaagttcat cccaggacgc tgccacacct acgaaggcga
     4321 caaagagtcc gcacagggcg gcataggcga ggcgatcgtc gacattcctg agattcctgg
     4381 gttcaaggac ttggagccca tggagcagtt catcgcacag gtcgatctgt gtgtggactg
     4441 cacaactggc tgcctcaaag ggcttgccaa cgtgcagtgt tctgacctgc tcaagaagtg
     4501 gctgccgcaa cgctgtgcga cctttgccag caagatccag ggccaggtgg acaagatcaa
     4561 gggggccggt ggtgactaaa ataaaatatc tttattttca ttacatctgt gtgttggttt
     4621 tttgtgtgcc cgggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg
     4681 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4741 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4801 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4861 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4921 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4981 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5041 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5101 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5161 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5221 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5281 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5341 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5401 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5461 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5521 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5581 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5641 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5701 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5761 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5821 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5881 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5941 atctggagcc ggtgagcgtg ggtctggcgg tatcattgca gcactggggc cagatggtaa
     6001 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6061 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6121 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6181 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6241 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6301 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6361 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6421 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6481 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6541 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6601 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6661 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6721 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6781 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6841 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6901 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6961 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7021 cgagtcagtg agcgaggaag cgga
//
