LOCUS       Exported                7778 bp DNA     circular SYN 07-MAR-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7778)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 7, 2021 from SnapGene 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7778
                     /organism="recombinant plasmid"
                     /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(1576..3150)
                     /codon_start=1
                     /label=oG(B19)
                     /translation="MVPQALLFVPLLVFPLCFGKFPIYTIPDKLGPWSPIDIHHLSCPN
                     NLVVEDEGCTNLSGFSYMELKVGYISAIKMNGFTCTGVVTEAETYTNFVGYVTTTFKRK
                     HFRPTPDACRAAYNWKMAGDPRYEESLHNPYPDYHWLRTVKTTKESLVIISPSVADLDP
                     YDRSLHSPVFPGGNCSGVAVSSTYCSTNHDYTIWMPENPRLGMSCDIFTNSRGKRASKG
                     SETCGFVDERGLYKSLKGACKLKLCGVLGLRLMDGTWVAMQTSNETKWCPPGQLVNLHD
                     FRSDEIEHLVVEELVKKREECLDALESIMTTKSVSFRRLSHLRKLVPGFGKAYTIFNKT
                     LMEADAHYKSVRTWNEIIPSKGCLRVGGRCHPHVNGVFFNGIILGPDGNVLIPEMQSSL
                     LQQHMELLVSSVIPLMHPLADPSTVFKNGDEAEDFVEVHLPDVHERISGVDLGLPNWGK
                     YVLLSAGALTALMLIIFLMTCWRRVNRSEPTQHNLRGTGREVSVTPQSGKIISSWESHK
                     SGGETRL"
     CDS             complement(3154..3210)
                     /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     3213..3246
                     /label=Fragment.FOR
     primer_bind     3228..3314
                     /label=Primer 1
     primer_bind     complement(3230..3316)
                     /label=Primer 2
     CDS             complement(3232..3297)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3301..3331)
                     /label=Fragment.REV
     CDS             complement(3319..3756)
                     /codon_start=1
                     /label=TVA
                     /translation="MARLLPALLLLLLPGNVTGNGSGNGFLSRCPPGQFRCSEPPGAHG
                     ECYPQDWLCDGHPDCDDGRDEWGCGTSAIPAVPTDNGTEAPTAPASGRALPARNHGRMW
                     MLITAGPLCCLVAVGGIAAWGKSKAKSRFDIFSLERASKGGG"
     protein_bind    complement(3763..3796)
                     /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(3847..3880)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3905..4493
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3972..3991)
                     /label=WHV5R
     CDS             complement(4376..4387)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4525..5001
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4585..4605)
                     /label=CCMV24
     primer_bind     complement(4589..4605)
                     /label=XL39rev
     repeat_region   5041..5181
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      5256..5711
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     5822..5841
                     /label=GEX3rev
     primer_bind     5851..5869
                     /label=GEX3rev25
     primer_bind     complement(5900..5918)
                     /label=QEfor
     promoter        5993..6097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6098..6958
                     /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(6143..6162)
                     /label=AmpStart
     primer_bind     6889..6908
                     /label=AmpStop
     rep_origin      7129..7717
                     /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 gcgccttaga gccgtgtctc gcccccagat ttatgacttt cccaactgct
     1621 gatgatttta ccgctctggg gtgtgacaga cacttctcta ccggtccccc tcagattgtg
     1681 ctgagttggc tcggagcggt tcactcgtcg ccagcaggtc atcaggaaaa tgatcagcat
     1741 gagagcagtc agtgctccag cggagagcag cacgtacttg ccccaattag gcagtccgag
     1801 atcgacgcca ctgatccgtt cgtgcacgtc gggcaggtgg acctccacaa aatcttcggc
     1861 ctcgtcgccg tttttgaaca ctgtgcttgg gtctgcgaga ggatgcatca ggggaatgac
     1921 gcttgacacg agcagttcca tgtgctgctg gagcagacta gactgcatct cggggatcag
     1981 cacgttacca tctggcccca gaatgatacc attaaagaag accccgttca catgtgggtg
     2041 acatcgaccc ccgactcgca ggcatccctt actaggaatg atttcattcc aggttctcac
     2101 agatttataa tgggcgtctg cctccatcag tgtcttgtta aagatggtgt atgcttttcc
     2161 gaagccaggg acgagcttcc tcaggtggct cagcctcctg aagctcacgc tcttggtagt
     2221 cataattgac tccagagcat cgagacattc ctcccttttc ttcacgagtt cctcgaccac
     2281 cagatgttcg atctcatcgc ttcggaagtc gtgcagattc acgagctgcc caggggggca
     2341 ccacttagtc tcgttagatg tctgcatagc gacccaggtt ccgtccatca gccggagtcc
     2401 cagcacgcca cagagtttca gcttgcaggc acctttgagg ctcttataca gccctctttc
     2461 atccacaaat ccacaagtct cgctgccttt tgaggccctc ttgcccctgc tgttggtgaa
     2521 gatgtcgcag gacatcccca gccttggatt ctcaggcatc cagattgtat aatcatggtt
     2581 ggtggaacag taagtggagc tgacggccac tccggagcag tttccgccgg gaaacactgg
     2641 actgtggaga gatcggtcgt aggggtccag gtcggcgaca gatggggaaa tgatgaccag
     2701 gctctctttt gtggtcttca cagtgcgcag ccaatgatag tctgggtaag gattgtgcag
     2761 gctttcctca tagcgggggt cgccagccat cttccagttg taagctgccc gacaggcgtc
     2821 tggtgttggc cgaaagtgtt ttctcttgaa agttgtggtg acatatccca caaaatttgt
     2881 gtaggtttct gcctcggtga ccacaccagt gcatgtgaac ccgttcatct taatagcaga
     2941 gatatagccc actttcagct ccatgtagga gaatccgctc aggttggtac agccttcgtc
     3001 ctcgaccacg agattgttag ggcatgacag atggtgaata tcgataggac tccaggggcc
     3061 cagcttgtcg gggatagtgt agatagggaa tttaccaaag cagagtggga agacgagcag
     3121 agggacgaag aggagagcct gtgggaccat ggttggcccg ggattctctt cgacatcccc
     3181 tgcttgtttc aacagggaga agttagtggc tccgcttccg ttgtcttcga acttgtcgtc
     3241 atcgtctttg tagtcgatat catgatcctt ataatcacca tcatgatcct tataatcgcc
     3301 ggtgctgccc ggttcgaacc cacccccctt ggatgccctt tcaagactga aaatgtcaaa
     3361 cctgcttttt gctttggact tcccccatgc agcgataccc cccacagcta ccaggcagca
     3421 caggggccct gcagtgatca gcatccacat gcggccgtga ttcctggctg gcaaagcacg
     3481 tccagaagca ggggcagtgg gagcctctgt gccgttgtcc gtgggcaccg cggggatcgc
     3541 gctggtcccg cacccccact cgtcccgccc gtcgtcgcag tcggggtgtc cgtcgcacag
     3601 ccagtcctgc gggtaacact ccccgtgggc accgggcggc tccgagcagc ggaactgacc
     3661 gggggggcaa cgggacaaaa aaccgttacc ggacccgtta ccggtcacgt taccgggcag
     3721 cagcagcagc agcagcgcgg gcagcagccg cgccatgcta gcataacttc gtataaagta
     3781 tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg
     3841 caaagaataa cttcgtataa tgtatgctat acgaagttat gaattcgata tcaagcttat
     3901 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3961 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4021 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4081 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     4141 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     4201 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4261 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     4321 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     4381 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     4441 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     4501 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     4561 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     4621 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     4681 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     4741 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     4801 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     4861 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     4921 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     4981 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     5041 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     5101 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     5161 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     5221 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     5281 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5341 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5401 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5461 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5521 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5581 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     5641 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5701 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5761 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5821 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5881 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5941 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6001 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6061 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6121 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6181 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6241 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6301 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6361 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6421 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6481 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6541 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6601 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6661 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6721 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6781 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6841 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6901 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6961 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7021 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7081 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7141 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7201 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7261 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7321 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7381 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7441 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7501 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7561 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7621 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7681 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7741 acggttcctg gccttttgct ggccttttgc tcacatgt
//
