LOCUS       Exported                8093 bp DNA     circular SYN 21-MAR-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8093)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 7, 2021 from SnapGene 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8093
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             5..136
                     /label=ITR
     protein_bind    216..249
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        284..1576
                     /label=CaMKII promoter
     regulatory      1592..1601
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1598..2035
                     /codon_start=1
                     /label=TVA
                     /translation="MARLLPALLLLLLPGNVTGNGSGNGFLSRCPPGQFRCSEPPGAHG
                     ECYPQDWLCDGHPDCDDGRDEWGCGTSAIPAVPTDNGTEAPTAPASGRALPARNHGRMW
                     MLITAGPLCCLVAVGGIAAWGKSKAKSRFDIFSLERASKGGG"
     CDS             2057..2122
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2144..2200
                     /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="ATNFSLLKQAGDVEENPGP"
     CDS             2204..3778
                     /codon_start=1
                     /label=N2cG
                     /translation="MVPQVLLFVLLLGFSLCFGKFPIYTIPDELGPWSPIDIHHLSCPN
                     NLVVEDEGCTNLSEFSYMELKVGYISAIKVNGFTCTGVVTEAETYTNFVGYVTTTFKRK
                     HFRPTPDACRAAYNWKMAGDPRYEESLHNPYPDYHWLRTVRTTKESLIIISPSVTDLDP
                     YDKSLHSRVFPGGKCSGITVSSTYCSTNHDYTIWMPENPRPRTPCDIFTNSRGKRASNG
                     NKTCGFVDERGLYKSLKGACRLKLCGVLGLRLMDGTWVAMQTSDETKWCPPDQLVNLHD
                     FRSDEIEHLVVEELVKKREECLDALESIMTTKSVSFRRLSHLRKLVPGFGKAYTIFNKT
                     LMEADAHYKSVRTWNEIIPSKGCLKVGGRCHPHVNGVFFNGIILGPDDHVLIPEMQSSL
                     LQQHMELLESSVIPLMHPLADPSTVFKEGDEAEDFVEVHLPDVYKQISGVDLGLPNWGK
                     YVLMTAGAMIGLVLIFSLMTWCRRANRPESKQRSFGGTGGNVSVTSQSGKVIPSWESYK
                     SGGEIRL"
     protein_bind    3819..3852
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3908..4496
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4379..4390)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4529..4753
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     LTR             4839..4970
                     /label=ITR
     promoter        complement(4974..5303)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4988..5123
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(5549..6137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6308..7168)
                     /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(7169..7273)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        8061..8079
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 ttctagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctaggt accttccagc aggtcgaggg acctaataac ttcgtatagc atacattata
      241 cgaagttata ttaagggttc caagcttaag cggccgcacg cgtttaacat tatggcctta
      301 ggtcacttca tctccatggg gttcttcttc tgattttcta gaaaatgaga tgggggtgca
      361 gagagcttcc tcagtgacct gcccagggtc acatcagaaa tgtcagagct agaacttgaa
      421 ctcagattac taatcttaaa ttccatgcct tgggggcatg caagtacgat atacagaagg
      481 agtgaactca ttagggcaga tgaccaatga gtttaggaaa gaagagtcca gggcagggta
      541 catctacacc acccgcccag ccctgggtga gtccagccac gttcacctca ttatagttgc
      601 ctctctccag tcctaccttg acgggaagca caagcagaaa ctgggacagg agccccagga
      661 gaccaaatct tcatggtccc tctgggagga tgggtgggga gagctgtggc agaggcctca
      721 ggaggggccc tgctgctcag tggtgacaga taggggtgag aaagcagaca gagtcattcc
      781 gtcagcattc tgggtctgtt tggtacttct tctcacgcta aggtggcggt gtgatatgca
      841 caatggctaa aaagcaggga gagctggaaa gaaacaagga cagagacaga ggccaagtca
      901 accagaccaa ttcccagagg aagcaaagaa accattacag agactacaag ggggaaggga
      961 aggagagatg aattagcttc ccctgtaaac cttagaaccc agctgttgcc agggcaacgg
     1021 ggcaatacct gtctcttcag aggagatgaa gttgccaggg taactacatc ctgtctttct
     1081 caaggaccat cccagaatgt ggcacccact agccgttacc atagcaactg cctctttgcc
     1141 ccacttaatc ccatcccgtc tgttaaaagg gccctatagt tggaggtggg ggaggtagga
     1201 agagcgatga tcacttgtgg actaagtttg ttcgcatccc cttctccaac cccctcagta
     1261 catcaccctg ggggaacagg gtccacttgc tcctgggccc acacagtcct gcagtattgt
     1321 gtatataagg ccagggcaaa gaggagcagg ttttaaagtg aaaggcaggc aggtgttggg
     1381 gaggcagtta ccggggcaac gggaacaggg cgtttcggag gtggttgcca tggggacctg
     1441 gatgctgacg aaggctcgcg aggctgtgag cagccacagt gccctgctca gaagccccaa
     1501 gctcgtcagt caagccggtt ctccgtttgc actcaggagc acgggcaggc gagtggcccc
     1561 tagttctggg ggcagcgggg gatccactag tgccaccatg gcgcggctgc tgcccgcgct
     1621 gctgctgctg ctgctgcccg gtaacgtgac cggtaacggg tccggtaacg gttttttgtc
     1681 ccgttgcccc cccggtcagt tccgctgctc ggagccgccc ggtgcccacg gggagtgtta
     1741 cccgcaggac tggctgtgcg acggacaccc cgactgcgac gacgggcggg acgagtgggg
     1801 gtgcgggacc agcgcgatcc ccgcggtgcc cacggacaac ggcacagagg ctcccactgc
     1861 ccctgcttct ggacgtgctt tgccagccag gaatcacggc cgcatgtgga tgctgatcac
     1921 tgcagggccc ctgtgctgcc tggtagctgt ggggggtatc gctgcatggg ggaagtccaa
     1981 agcaaaaagc aggtttgaca ttttcagtct tgaaagggca tccaaggggg gtgggttcga
     2041 accgggcagc accggcgatt ataaggatca tgatggtgat tataaggatc atgatatcga
     2101 ctacaaagac gatgacgaca agttcgaaga caacggaagc ggagccacta acttctccct
     2161 gttgaaacaa gcaggggatg tcgaagagaa tcccgggcca accatggttc ctcaggttct
     2221 tttgtttgta ctccttctgg gtttttcgtt gtgtttcggg aagttcccca tttacacgat
     2281 accagacgaa cttggtccct ggagccctat tgacatacac catctcagct gtccaaataa
     2341 cctggttgtg gaggatgaag gatgtaccaa cctgtccgag ttctcctaca tggaactcaa
     2401 agtgggatac atctcagcca tcaaagtgaa cgggttcact tgcacaggtg ttgtgacaga
     2461 ggcagagacc tacaccaact ttgttggtta tgtcacaacc acattcaaga gaaagcattt
     2521 ccgccccacc ccagacgcat gtagagccgc gtataactgg aagatggccg gtgaccccag
     2581 atatgaagag tccctacaca atccataccc cgactaccac tggcttcgaa ctgtaagaac
     2641 caccaaagag tccctcatta tcatatcccc aagtgtgaca gatttggacc catatgacaa
     2701 atcccttcac tcaagggtct tccctggcgg aaagtgctca ggaataacgg tgtcctctac
     2761 ctactgctca actaaccatg attacaccat ttggatgccc gagaatccga gaccaaggac
     2821 accttgtgac atttttacca atagcagagg gaagagagca tccaacggga acaagacttg
     2881 cggctttgtg gatgaaagag gcctgtataa gtctctaaaa ggagcatgca ggctcaagtt
     2941 atgtggagtt cttggactta gacttatgga tggaacatgg gtcgcgatgc aaacatcaga
     3001 tgagaccaaa tggtgccctc cagatcagtt ggtgaatttg cacgactttc gctcagacga
     3061 gattgagcat ctcgttgtgg aggagttagt caagaaaaga gaggaatgtc tggatgcatt
     3121 agagtccatc atgaccacca agtcagtaag tttcagacgt ctcagtcacc tgagaaaact
     3181 tgtcccaggg tttggaaaag catataccat attcaacaaa accttgatgg aggctgatgc
     3241 tcactacaag tcagtccgga cctggaatga gatcatcccc tcaaaagggt gtttgaaagt
     3301 tggaggaagg tgccatcctc atgtgaacgg ggtgtttttc aatggtataa tattagggcc
     3361 tgacgaccat gtcctaatcc cagagatgca atcatccctc ctccagcaac atatggagtt
     3421 gttggaatct tcagttatcc ccctgatgca ccccctggca gacccttcta cagttttcaa
     3481 agaaggtgat gaggctgagg attttgttga agttcacctc cccgatgtgt acaaacagat
     3541 ctcaggggtt gacctgggtc tcccgaactg gggaaagtat gtattgatga ctgcaggggc
     3601 catgattggc ctggtgttga tattttccct aatgacatgg tgcagaagag ccaatcgacc
     3661 agaatcgaaa caacgcagtt ttggagggac aggggggaat gtgtcagtca cttcccaaag
     3721 cggaaaagtc ataccttcat gggaatcata taagagtgga ggtgagatca gactgtgaga
     3781 tgtacaagta aggcgcgccg aattcgtcga gggacctaat aacttcgtat agcatacatt
     3841 atacgaagtt atacatgttt aagggttccg gttccactag gtacaattcg atatcaagct
     3901 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3961 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4021 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4081 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4141 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4201 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4261 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     4321 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4381 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4441 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     4501 ataccgtcga ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4561 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4621 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4681 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4741 gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgactag agcatggcta
     4801 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     4861 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4921 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagag ctttttgcaa
     4981 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg
     5041 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     5101 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     5161 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     5221 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     5281 tttccacacc ctaactgaca cacattccac agctgcatta atgaatcggc caacgcgcgg
     5341 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5401 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5461 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5521 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5581 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5641 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5701 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5761 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5821 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5881 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5941 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6001 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6061 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6121 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6181 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6241 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6301 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6361 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6421 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6481 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6541 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6601 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6661 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6721 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6781 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6841 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6901 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6961 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7021 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7081 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7141 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7201 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7261 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     7321 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     7381 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     7441 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     7501 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cattcgacgc
     7561 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     7621 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     7681 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     7741 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     7801 atgccggcca cgatgcgtcc ggcgtagagg atctggctag cgatgaccct gctgattggt
     7861 tcgctgacca tttccgggtg cgggacggcg ttaccagaaa ctcagaaggt tcgtccaacc
     7921 aaaccgactc tgacggcagt ttacgagaga gatgataggg tctgcttcag taagccagat
     7981 gctacacaat taggcttgta catattgtcg ttagaacgcg gctacaatta atacataacc
     8041 ttatgtatca tacacatacg atttaggtga cactatagaa tacacggaat taa
//
