LOCUS       Exported                7823 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 7823)
  AUTHORS   Yoav Ben Simon
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 7, 2021 from SnapGene 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7823
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     promoter        156..1448
                     /label=CaMKII promoter
     protein_bind    1476..1509
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     protein_bind    1560..1593
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1615..3189)
                     /codon_start=1
                     /label=N2cG
                     /translation="MVPQVLLFVLLLGFSLCFGKFPIYTIPDELGPWSPIDIHHLSCPN
                     NLVVEDEGCTNLSEFSYMELKVGYISAIKVNGFTCTGVVTEAETYTNFVGYVTTTFKRK
                     HFRPTPDACRAAYNWKMAGDPRYEESLHNPYPDYHWLRTVRTTKESLIIISPSVTDLDP
                     YDKSLHSRVFPGGKCSGITVSSTYCSTNHDYTIWMPENPRPRTPCDIFTNSRGKRASNG
                     NKTCGFVDERGLYKSLKGACRLKLCGVLGLRLMDGTWVAMQTSDETKWCPPDQLVNLHD
                     FRSDEIEHLVVEELVKKREECLDALESIMTTKSVSFRRLSHLRKLVPGFGKAYTIFNKT
                     LMEADAHYKSVRTWNEIIPSKGCLKVGGRCHPHVNGVFFNGIILGPDDHVLIPEMQSSL
                     LQQHMELLESSVIPLMHPLADPSTVFKEGDEAEDFVEVHLPDVYKQISGVDLGLPNWGK
                     YVLMTAGAMIGLVLIFSLMTWCRRANRPESKQRSFGGTGGNVSVTSQSGKVIPSWESYK
                     SGGEIRL"
     CDS             complement(3193..3249)
                     /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             complement(3271..3336)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3358..3795)
                     /codon_start=1
                     /label=TVA
                     /translation="MARLLPALLLLLLPGNVTGNGSGNGFLSRCPPGQFRCSEPPGAHG
                     ECYPQDWLCDGHPDCDDGRDEWGCGTSAIPAVPTDNGTEAPTAPASGRALPARNHGRMW
                     MLITAGPLCCLVAVGGIAAWGKSKAKSRFDIFSLERASKGGG"
     regulatory      3792..3801
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(3808..3841)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     protein_bind    complement(3892..3925)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3950..4538
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4421..4432)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4570..5046
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   5086..5226
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      5301..5756
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6038..6142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6143..7003
                     /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"
     rep_origin      7174..7762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttaac attatggcct taggtcactt
      181 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt
      241 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt
      301 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact
      361 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca
      421 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc
      481 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat
      541 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc
      601 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat
      661 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct
      721 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc
      781 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga
      841 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac
      901 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc
      961 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa
     1021 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat
     1081 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc
     1141 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa
     1201 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt
     1261 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga
     1321 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca
     1381 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg
     1441 ggggcagcgg gggatcctct agagtcgact ccggaataac ttcgtatagg atactttata
     1501 cgaagttatg cagaatggta gctggattgt agctgctatt agcaatatga aacctcttaa
     1561 taacttcgta tagcatacat tatacgaagt tatggcgcgc cttacttgta catctcacag
     1621 tctgatctca cctccactct tatatgattc ccatgaaggt atgacttttc cgctttggga
     1681 agtgactgac acattccccc ctgtccctcc aaaactgcgt tgtttcgatt ctggtcgatt
     1741 ggctcttctg caccatgtca ttagggaaaa tatcaacacc aggccaatca tggcccctgc
     1801 agtcatcaat acatactttc cccagttcgg gagacccagg tcaacccctg agatctgttt
     1861 gtacacatcg gggaggtgaa cttcaacaaa atcctcagcc tcatcacctt ctttgaaaac
     1921 tgtagaaggg tctgccaggg ggtgcatcag ggggataact gaagattcca acaactccat
     1981 atgttgctgg aggagggatg attgcatctc tgggattagg acatggtcgt caggccctaa
     2041 tattatacca ttgaaaaaca ccccgttcac atgaggatgg caccttcctc caactttcaa
     2101 acaccctttt gaggggatga tctcattcca ggtccggact gacttgtagt gagcatcagc
     2161 ctccatcaag gttttgttga atatggtata tgcttttcca aaccctggga caagttttct
     2221 caggtgactg agacgtctga aacttactga cttggtggtc atgatggact ctaatgcatc
     2281 cagacattcc tctcttttct tgactaactc ctccacaacg agatgctcaa tctcgtctga
     2341 gcgaaagtcg tgcaaattca ccaactgatc tggagggcac catttggtct catctgatgt
     2401 ttgcatcgcg acccatgttc catccataag tctaagtcca agaactccac ataacttgag
     2461 cctgcatgct ccttttagag acttatacag gcctctttca tccacaaagc cgcaagtctt
     2521 gttcccgttg gatgctctct tccctctgct attggtaaaa atgtcacaag gtgtccttgg
     2581 tctcggattc tcgggcatcc aaatggtgta atcatggtta gttgagcagt aggtagagga
     2641 caccgttatt cctgagcact ttccgccagg gaagaccctt gagtgaaggg atttgtcata
     2701 tgggtccaaa tctgtcacac ttggggatat gataatgagg gactctttgg tggttcttac
     2761 agttcgaagc cagtggtagt cggggtatgg attgtgtagg gactcttcat atctggggtc
     2821 accggccatc ttccagttat acgcggctct acatgcgtct ggggtggggc ggaaatgctt
     2881 tctcttgaat gtggttgtga cataaccaac aaagttggtg taggtctctg cctctgtcac
     2941 aacacctgtg caagtgaacc cgttcacttt gatggctgag atgtatccca ctttgagttc
     3001 catgtaggag aactcggaca ggttggtaca tccttcatcc tccacaacca ggttatttgg
     3061 acagctgaga tggtgtatgt caatagggct ccagggacca agttcgtctg gtatcgtgta
     3121 aatggggaac ttcccgaaac acaacgaaaa acccagaagg agtacaaaca aaagaacctg
     3181 aggaaccatg gttggcccgg gattctcttc gacatcccct gcttgtttca acagggagaa
     3241 gttagtggct ccgcttccgt tgtcttcgaa cttgtcgtca tcgtctttgt agtcgatatc
     3301 atgatcctta taatcaccat catgatcctt ataatcgccg gtgctgcccg gttcgaaccc
     3361 accccccttg gatgcccttt caagactgaa aatgtcaaac ctgctttttg ctttggactt
     3421 cccccatgca gcgatacccc ccacagctac caggcagcac aggggccctg cagtgatcag
     3481 catccacatg cggccgtgat tcctggctgg caaagcacgt ccagaagcag gggcagtggg
     3541 agcctctgtg ccgttgtccg tgggcaccgc ggggatcgcg ctggtcccgc acccccactc
     3601 gtcccgcccg tcgtcgcagt cggggtgtcc gtcgcacagc cagtcctgcg ggtaacactc
     3661 cccgtgggca ccgggcggct ccgagcagcg gaactgaccg ggggggcaac gggacaaaaa
     3721 accgttaccg gacccgttac cggtcacgtt accgggcagc agcagcagca gcagcgcggg
     3781 cagcagccgc gccatggtgg cactagcata acttcgtata aagtatccta tacgaagtta
     3841 tttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag aataacttcg
     3901 tataatgtat gctatacgaa gttatgaatt cgatatcaag cttatcgata atcaacctct
     3961 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4021 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4081 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     4141 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     4201 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     4261 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     4321 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     4381 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     4441 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     4501 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     4561 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     4621 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     4681 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     4741 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     4801 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     4861 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     4921 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     4981 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     5041 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     5101 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     5161 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     5221 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5281 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5341 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5401 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5461 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     5521 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5581 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5641 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5701 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     5761 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     5821 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5881 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5941 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6001 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6061 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6121 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6181 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6241 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6301 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6361 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6421 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6481 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6541 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6601 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6661 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6721 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6781 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6841 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6901 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6961 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7021 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7081 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7141 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7201 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7261 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7321 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7381 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7441 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7501 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7561 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7621 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7681 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7741 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7801 ttgctggcct tttgctcaca tgt
//
