LOCUS       Exported                6931 bp DNA     circular SYN 01-MAR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6931)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 7, 2021 from SnapGene 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6931
                     /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        163..699
                     /label=mDLX promoter
     intron          860..992
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1034..1043
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1499..1564
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1586..1642
                     /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"
     misc_feature    3277..3865
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3748..3759)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4034..4115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   4194..4334
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4409..4864
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5146..5250
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5251..6111
                     /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      6282..6870
                     /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 cgcgtttaat taagaattct atacactcac
      181 agtggtttgg catatatttg gtgaaatttt ttaaggaaaa attagtgttg gtttcgatat
      241 atggtagctt tttctctaac ataatttgaa taattcagca aagccctact accagctgta
      301 cttctgcagc ctcttccatt ctttccagca ttataatttt ggttaatttt caattttagg
      361 tcctacgtct ctgcaatttg tgtatgaata acagaataat ttccctcttt tgtttcgcct
      421 ttcctgttcc tgaatctaaa taaagatggc tttttagtat taaaagtgga agaaaattac
      481 aggtaattat ctttgacggt aaaaacgctg taatcagcgg gctacatgaa aaattactct
      541 aattatggct gcatttaaga gaatggaaaa aaaccttctt gtggataaaa accttaaatt
      601 gtccccaatg tctgcttcaa attggatggc actgcagctg gaggctttgt tcagaattga
      661 tcctggggag ctacgaaccc aaagtttcac agtaggaagc ttgatatcga attcctgcag
      721 cccgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctagcct
      781 gcaggtcgag gagcgcagcc ttccagaagc agagcgcggc gccttaagct gcagaagttg
      841 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
      901 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
      961 tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc
     1021 ttaagaaact agtgccacca tggcgcggct gctgcccgcg ctgctgctgc tgctgctgcc
     1081 cggtaacgtg accggtaacg ggtccggtaa cggttttttg tcccgttgcc cccccggtca
     1141 gttccgctgc tcggagccgc ccggtgccca cggggagtgt tacccgcagg actggctgtg
     1201 cgacggacac cccgactgcg acgacgggcg ggacgagtgg gggtgcggga ccagcgcgat
     1261 ccccgcggtg cccacggaca acggcacaga ggctcccact gcccctgctt ctggacgtgc
     1321 tttgccagcc aggaatcacg gccgcatgtg gatgctgatc actgcagggc ccctgtgctg
     1381 cctggtagct gtggggggta tcgctgcatg ggggaagtcc aaagcaaaaa gcaggtttga
     1441 cattttcagt cttgaaaggg catccaaggg gggtgggttc gaaccgggca gcaccggcga
     1501 ttataaggat catgatggtg attataagga tcatgatatc gactacaaag acgatgacga
     1561 caagttcgaa gacaacggaa gcggagccac taacttctcc ctgttgaaac aagcagggga
     1621 tgtcgaagag aatcccgggc caaccatggt tcctcaggtt cttttgtttg tactccttct
     1681 gggtttttcg ttgtgtttcg ggaagttccc catttacacg ataccagacg aacttggtcc
     1741 ctggagccct attgacatac accatctcag ctgtccaaat aacctggttg tggaggatga
     1801 aggatgtacc aacctgtccg agttctccta catggaactc aaagtgggat acatctcagc
     1861 catcaaagtg aacgggttca cttgcacagg tgttgtgaca gaggcagaga cctacaccaa
     1921 ctttgttggt tatgtcacaa ccacattcaa gagaaagcat ttccgcccca ccccagacgc
     1981 atgtagagcc gcgtataact ggaagatggc cggtgacccc agatatgaag agtccctaca
     2041 caatccatac cccgactacc actggcttcg aactgtaaga accaccaaag agtccctcat
     2101 tatcatatcc ccaagtgtga cagatttgga cccatatgac aaatcccttc actcaagggt
     2161 cttccctggc ggaaagtgct caggaataac ggtgtcctct acctactgct caactaacca
     2221 tgattacacc atttggatgc ccgagaatcc gagaccaagg acaccttgtg acatttttac
     2281 caatagcaga gggaagagag catccaacgg gaacaagact tgcggctttg tggatgaaag
     2341 aggcctgtat aagtctctaa aaggagcatg caggctcaag ttatgtggag ttcttggact
     2401 tagacttatg gatggaacat gggtcgcgat gcaaacatca gatgagacca aatggtgccc
     2461 tccagatcag ttggtgaatt tgcacgactt tcgctcagac gagattgagc atctcgttgt
     2521 ggaggagtta gtcaagaaaa gagaggaatg tctggatgca ttagagtcca tcatgaccac
     2581 caagtcagta agtttcagac gtctcagtca cctgagaaaa cttgtcccag ggtttggaaa
     2641 agcatatacc atattcaaca aaaccttgat ggaggctgat gctcactaca agtcagtccg
     2701 gacctggaat gagatcatcc cctcaaaagg gtgtttgaaa gttggaggaa ggtgccatcc
     2761 tcatgtgaac ggggtgtttt tcaatggtat aatattaggg cctgacgacc atgtcctaat
     2821 cccagagatg caatcatccc tcctccagca acatatggag ttgttggaat cttcagttat
     2881 ccccctgatg caccccctgg cagacccttc tacagttttc aaagaaggtg atgaggctga
     2941 ggattttgtt gaagttcacc tccccgatgt gtacaaacag atctcagggg ttgacctggg
     3001 tctcccgaac tggggaaagt atgtattgat gactgcaggg gccatgattg gcctggtgtt
     3061 gatattttcc ctaatgacat ggtgcagaag agccaatcga ccagaatcga aacaacgcag
     3121 ttttggaggg acagggggga atgtgtcagt cacttcccaa agcggaaaag tcataccttc
     3181 atgggaatca tataagagtg gaggtgagat cagactgtga gatgtacaag taaggcgcgc
     3241 cacccctgca gggaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     3301 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3361 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3421 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3481 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3541 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3601 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3661 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     3721 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3781 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3841 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctgtgata
     3901 gcggccatca agctggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     3961 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4021 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4081 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4141 catcaatgta tcagcttatc gataccgcat gcacgtgcgg accgagcggc cgcaggaacc
     4201 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4261 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4321 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4381 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4441 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4501 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4561 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4621 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4681 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4741 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4801 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4861 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4921 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4981 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5041 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5101 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5161 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5221 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5281 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5341 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5401 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5461 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5521 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5581 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5641 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5701 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5761 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5821 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5881 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5941 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6001 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6061 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6121 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6181 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6241 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6301 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6361 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6421 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6481 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6541 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6601 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6661 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6721 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6781 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6841 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6901 ctggcctttt gctggccttt tgctcacatg t
//
