LOCUS       Exported                7797 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 7797)
  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..7797
                     /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        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1alpha
     protein_bind    1450..1483
                     /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    1534..1567
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1589..3160)
                     /codon_start=1
                     /label=N2C-G
                     /translation="VPQVLLFVLLLGFSLCFGKFPIYTIPDELGPWSPIDIHHLSCPNN
                     LVVEDEGCTNLSEFSYMELKVGYISAIKVNGFTCTGVVTEAETYTNFVGYVTTTFKRKH
                     FRPTPDACRAAYNWKMAGDPRYEESLHNPYPDYHWLRTVRTTKESLIIISPSVTDLDPY
                     DKSLHSRVFPGGKCSGITVSSTYCSTNHDYTIWMPENPRPRTPCDIFTNSRGKRASNGN
                     KTCGFVDERGLYKSLKGACRLKLCGVLGLRLMDGTWVAMQTSDETKWCPPDQLVNLHDF
                     RSDEIEHLVVEELVKKREECLDALESIMTTKSVSFRRLSHLRKLVPGFGKAYTIFNKTL
                     MEADAHYKSVRTWNEIIPSKGCLKVGGRCHPHVNGVFFNGIILGPDDHVLIPEMQSSLL
                     QQHMELLESSVIPLMHPLADPSTVFKEGDEAEDFVEVHLPDVYKQISGVDLGLPNWGKY
                     VLMTAGAMIGLVLIFSLMTWCRRANRPESKQRSFGGTGGNVSVTSQSGKVIPSWESYKS
                     GGEIRL"
     CDS             complement(3161..3163)
                     /codon_start=1
                     /label=oG(B19)
                     /translation="M"
     CDS             complement(3167..3223)
                     /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"
     primer_bind     3226..3259
                     /label=Fragment.FOR
     primer_bind     3241..3327
                     /label=Primer 1
     primer_bind     complement(3243..3329)
                     /label=Primer 2
     CDS             complement(3245..3310)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3314..3344)
                     /label=Fragment.REV
     CDS             complement(3332..3769)
                     /codon_start=1
                     /label=TVA
                     /translation="MARLLPALLLLLLPGNVTGNGSGNGFLSRCPPGQFRCSEPPGAHG
                     ECYPQDWLCDGHPDCDDGRDEWGCGTSAIPAVPTDNGTEAPTAPASGRALPARNHGRMW
                     MLITAGPLCCLVAVGGIAAWGKSKAKSRFDIFSLERASKGGG"
     regulatory      3770..3775
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(3782..3815)
                     /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(3866..3899)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3924..4512
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3991..4010)
                     /label=WHV5R
     primer_bind     4279..4291
                     /label=CCMV24
     CDS             complement(4395..4406)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4544..5020
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4604..4624)
                     /label=CCMV24
     primer_bind     complement(4608..4624)
                     /label=XL39rev
     repeat_region   5060..5200
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      5275..5730
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     5841..5860
                     /label=GEX3rev
     primer_bind     5870..5888
                     /label=GEX3rev25
     primer_bind     complement(5919..5937)
                     /label=QEfor
     promoter        6012..6116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6117..6977
                     /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"
     primer_bind     complement(6162..6181)
                     /label=AmpStart
     primer_bind     6908..6927
                     /label=AmpStop
     rep_origin      7148..7736
                     /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 cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1561 aagttatggc gcgccttact tgtacatctc acagtctgat ctcacctcca ctcttatatg
     1621 attcccatga aggtatgact tttccgcttt gggaagtgac tgacacattc ccccctgtcc
     1681 ctccaaaact gcgttgtttc gattctggtc gattggctct tctgcaccat gtcattaggg
     1741 aaaatatcaa caccaggcca atcatggccc ctgcagtcat caatacatac tttccccagt
     1801 tcgggagacc caggtcaacc cctgagatct gtttgtacac atcggggagg tgaacttcaa
     1861 caaaatcctc agcctcatca ccttctttga aaactgtaga agggtctgcc agggggtgca
     1921 tcagggggat aactgaagat tccaacaact ccatatgttg ctggaggagg gatgattgca
     1981 tctctgggat taggacatgg tcgtcaggcc ctaatattat accattgaaa aacaccccgt
     2041 tcacatgagg atggcacctt cctccaactt tcaaacaccc ttttgagggg atgatctcat
     2101 tccaggtccg gactgacttg tagtgagcat cagcctccat caaggttttg ttgaatatgg
     2161 tatatgcttt tccaaaccct gggacaagtt ttctcaggtg actgagacgt ctgaaactta
     2221 ctgacttggt ggtcatgatg gactctaatg catccagaca ttcctctctt ttcttgacta
     2281 actcctccac aacgagatgc tcaatctcgt ctgagcgaaa gtcgtgcaaa ttcaccaact
     2341 gatctggagg gcaccatttg gtctcatctg atgtttgcat cgcgacccat gttccatcca
     2401 taagtctaag tccaagaact ccacataact tgagcctgca tgctcctttt agagacttat
     2461 acaggcctct ttcatccaca aagccgcaag tcttgttccc gttggatgct ctcttccctc
     2521 tgctattggt aaaaatgtca caaggtgtcc ttggtctcgg attctcgggc atccaaatgg
     2581 tgtaatcatg gttagttgag cagtaggtag aggacaccgt tattcctgag cactttccgc
     2641 cagggaagac ccttgagtga agggatttgt catatgggtc caaatctgtc acacttgggg
     2701 atatgataat gagggactct ttggtggttc ttacagttcg aagccagtgg tagtcggggt
     2761 atggattgtg tagggactct tcatatctgg ggtcaccggc catcttccag ttatacgcgg
     2821 ctctacatgc gtctggggtg gggcggaaat gctttctctt gaatgtggtt gtgacataac
     2881 caacaaagtt ggtgtaggtc tctgcctctg tcacaacacc tgtgcaagtg aacccgttca
     2941 ctttgatggc tgagatgtat cccactttga gttccatgta ggagaactcg gacaggttgg
     3001 tacatccttc atcctccaca accaggttat ttggacagct gagatggtgt atgtcaatag
     3061 ggctccaggg accaagttcg tctggtatcg tgtaaatggg gaacttcccg aaacacaacg
     3121 aaaaacccag aaggagtaca aacaaaagaa cctgaggaac catggttggc ccgggattct
     3181 cttcgacatc ccctgcttgt ttcaacaggg agaagttagt ggctccgctt ccgttgtctt
     3241 cgaacttgtc gtcatcgtct ttgtagtcga tatcatgatc cttataatca ccatcatgat
     3301 ccttataatc gccggtgctg cccggttcga acccaccccc cttggatgcc ctttcaagac
     3361 tgaaaatgtc aaacctgctt tttgctttgg acttccccca tgcagcgata ccccccacag
     3421 ctaccaggca gcacaggggc cctgcagtga tcagcatcca catgcggccg tgattcctgg
     3481 ctggcaaagc acgtccagaa gcaggggcag tgggagcctc tgtgccgttg tccgtgggca
     3541 ccgcggggat cgcgctggtc ccgcaccccc actcgtcccg cccgtcgtcg cagtcggggt
     3601 gtccgtcgca cagccagtcc tgcgggtaac actccccgtg ggcaccgggc ggctccgagc
     3661 agcggaactg accggggggg caacgggaca aaaaaccgtt accggacccg ttaccggtca
     3721 cgttaccggg cagcagcagc agcagcagcg cgggcagcag ccgcgccatg gtggcactag
     3781 cataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga aattatcact
     3841 gttattcttt agaatggtgc aaagaataac ttcgtataat gtatgctata cgaagttatg
     3901 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     3961 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4021 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4081 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4141 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4201 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4261 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     4321 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     4381 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4441 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4501 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga
     4561 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     4621 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     4681 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     4741 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     4801 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     4861 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     4921 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     4981 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     5041 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     5101 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     5161 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     5221 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     5281 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5341 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5401 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5461 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     5521 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5581 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     5641 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5701 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     5761 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5821 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5881 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5941 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6001 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6061 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6121 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6181 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6241 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6301 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6361 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6421 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6481 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6541 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6601 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6661 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6721 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6781 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6841 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6901 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6961 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7021 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7081 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7141 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7201 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7261 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     7321 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7381 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7441 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7501 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7561 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7621 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7681 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7741 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
