LOCUS       Exported                7124 bp ds-DNA     circular SYN 27-MAR-2019
DEFINITION  synthetic circular DNA
ACCESSION   VB181016-1178faa
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7124)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported to SnapGene on Oct 16, 2018
REFERENCE   2  (bases 1 to 7124)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 15, 2021 from SnapGene 5.1.7
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Exported to
            SnapGene on Oct 16, 2018"
FEATURES             Location/Qualifiers
     source          1..7124
                     /organism="synthetic DNA construct"
     misc_feature    1..141
                     /label=5' ITR
                     /label=nonstandard type:  ITR
                     /note="/application_notes=Allows replication of the viral 
                     genomeand its packaging into virus."
                     /note="/description=AAV 5' inverted terminal repeat 
                     (functionalequivalent of wild-type 5' ITR)"
                     /note="/official_designation=5' ITR"
                     /note="/uuid=5f14ec1a645a407dab2725d71bd1c86a"
     promoter        169..1647
                     /label={Prk2r}
                     /note="/application_notes="
                     /note="/official_designation={Prk2r}"
                     /note="/uuid=ab32d9e901dc437789cda2f9d8f09f0e"
     misc_feature    1672..1677
                     /label=Kozak
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Facilitates translation 
                     initiation ofATG start codon downstream of the Kozak 
                     sequence."
                     /note="/description=Kozak translation initiation sequence"
                     /note="/full_name=Kozak"
                     /note="/official_designation=Kozak"
                     /note="/uuid=5f18b25a1234443494a618363f09f052"
     misc_feature    1678..2730
                     /label=Cre(ns)
                     /label=nonstandard type:  ORF
                     /note="/application_notes=Most commonly used 
                     site-specificrecombinase"
                     /note="/description=Cyclization recombinase with SV40 
                     nuclearlocalization signal"
                     /note="/full_name=Cyclization recombinase(ns)"
                     /note="/official_designation=Cre(ns)"
                     /note="/uuid=9cd887825af842d5a920485ce7b2ceba"
     misc_feature    2731..2793
                     /label=T2A
                     /label=nonstandard type:  Linker
                     /note="/application_notes=Causes co-translational cleavage 
                     of theencoded polypeptide. Multiple proteins can be made 
                     from a polycistronic transcript containing multiple ORFs 
                     separated by 2A. P2A and T2A have higher cleavage 
                     efficiency compared to other 2As."
                     /note="/description=Self-cleaving 2A peptide from thosea 
                     asignavirus"
                     /note="/full_name=Thosea asigna virus 2A peptide"
                     /note="/official_designation=T2A"
                     /note="/uuid=995274d0c39f4cbdbab66fb78aa938d6"
     misc_feature    2794..3513
                     /label=Venus
                     /label=nonstandard type:  ORF
                     /note="/application_notes=Fast maturation and high 
                     tolerance toacidosis and Cl- compared to its predecessor 
                     YFP."
                     /note="/description=Variant of yellow fluorescent protein 
                     (YFP)generated by mutagenesis; YFP is itself a variant of 
                     EGFP generated by mutagenesis"
                     /note="/full_name=Venus"
                     /note="/official_designation=Venus"
                     /note="/uuid=0fe2a74c66e4407b88d6aa5c10887f0c"
     misc_feature    3544..4141
                     /label=WPRE
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Enhances virus stability in 
                     packagingcells, leading to higher titer of packaged virus; 
                     enhances higher expression of transgenes."
                     /note="/description=Woodchuck hepatitis virus 
                     posttranscriptionalregulatory element"
                     /note="/full_name=Woodchuck hepatitis virus 
                     posttranscriptionalregulatory element"
                     /note="/official_designation=WPRE"
                     /note="/uuid=0aedff8308474b369c622433538d7836"
     polyA_signal    4172..4379
                     /label=BGH pA
                     /note="/application_notes=Allows transcription termination 
                     andpolyadenylation of mRNA transcribed by Pol II RNA 
                     polymerase."
                     /note="/description=Bovine growth hormone polyadenylation 
                     signal"
                     /note="/full_name=Bovine growth hormone polyadenylation"
                     /note="/official_designation=BGH pA"
                     /note="/uuid=4a09734ca6c745b3aadc757c1638b651"
     misc_feature    4387..4527
                     /label=3' ITR
                     /label=nonstandard type:  ITR
                     /note="/application_notes=Allows replication of the viral 
                     genomeand its packaging into virus."
                     /note="/description=AAV 3' inverted terminal repeat"
                     /note="/official_designation=3' ITR"
                     /note="/uuid=5b8101593b634e64aa9eef9434216f71"
     misc_feature    5444..6304
                     /label=Ampicillin
                     /label=nonstandard type:  ORF
                     /note="/application_notes=Allows E. coli to be resistant 
                     toampicillin."
                     /note="/description=Ampicillin resistance gene"
                     /note="/full_name=Ampicillin resistance gene"
                     /note="/official_designation=Ampicillin"
                     /note="/uuid=5bcc09480c0e4c07b95ddccd8242d769"
     rep_origin      6475..7063
                     /direction=RIGHT
                     /label=pUC ori
                     /note="/application_notes=Facilitates plasmid replication 
                     in E.coli; regulates high-copy plasmid number (500-700)."
                     /note="/description=pUC origin of replication"
                     /note="/full_name=pUC origin of replication"
                     /note="/official_designation=pUC ori"
                     /note="/uuid=b978ad4ff785460db448caf63b728237"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttctagacaa ctttgtatag aaaagttgct gcacacttag
      181 tggaagccac aaatgcttgc agaggacagt gctccgtctc aaggatggcc agctgctgag
      241 gtcacaacgg gttcaagagc ttacaaaacc cttggaagga ccacccggaa gtctgagacc
      301 ctaattcagt agatctagga ggcaggcggg tctcgggggg cggagtcggg cgccgggagg
      361 ggcgggacac ctgggctggg tccggggctg ttggatgagc tgcagagcga agcagcaact
      421 ccagaaagac tgatgcgagg agagcaggac caagcgacac acgcccacca agtaggcaag
      481 cctcaaccag agctagtgtt cccagagccc gggccgcgcc cacctatggt ggcgccaccg
      541 gccagggagt gcgtccctga aagcaccgag gcaagtgagt accgggttgc ggcgggcggg
      601 cggacttggg ggcactccgc gtctgttcca ggagatcacc tcctacctcc gaaccctggg
      661 attggggact aggggactgg agtggagccc cgaaatcgcg gactcagtgg aaggcgtacg
      721 gacaagggtg ggggtatctg cgtgtttcct gcgccctgca ggcacctctg gtgcccggct
      781 ttgtttcttg agcttaaatt ccccactggg tacctaatcc tgtaccagtg ttgcactgag
      841 tgagtgagag acgtgggaac cttcaaacgg gctccctctt cccacagtct acgggctggt
      901 gatgagaagg cggctggaac tcccgcttat tggtccccgg tggcgatctt tgggagacca
      961 atagacgccc cagagggagg acactgggat ccagactcca ctggaacccc gcttttcagg
     1021 tagggtggag cgtgcggctg attcctgtgt agcaaatcgg ggtttcacta acgccggcag
     1081 agtggcgatt cctgtccccc agcgctctcc ctcgtaaaac tgggcagacc tcgctatcca
     1141 gtttgatagc ttcttgctac ccttacccgt tgctcccgta gctcccgcac tggctggtgt
     1201 gctgaggtgg gagaggaagg cagttcattg agacagacta agtcaggctg gagtcctctc
     1261 tccagcccct cgcttgcccc atagcagcta cctcctggga gctagcattt tggagactct
     1321 attttgtctc ccttccggct aagggtgcat actcttcttg ggtttctctg aagattccag
     1381 ttcattctgt gaccagcaag cagacggcct cccgtcttct tggccctggg tgactctggt
     1441 tggactgtac ctagctaact cttgggggga ctagccagat ctgatgtttg ctgaccagtg
     1501 gttttatctg cccccagatc ctggatggta tctgttctcc ctaaggattc taacagggac
     1561 ctgcactcac tgaccccagc agaactgctg aactccacgt gagcgcatct ccctgataca
     1621 caccagccca cctgtagcat catcaaccaa gtttgtacaa aaaagcaggc tgccaccatg
     1681 ggcccaaaga agaagagaaa ggtttcgaat ttactgaccg tacaccaaaa tttgcctgca
     1741 ttaccggtcg atgcaacgag tgatgaggtt cgcaagaacc tgatggacat gttcagggat
     1801 cgccaggcgt tttctgagca tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg
     1861 gcatggtgca agttgaataa ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat
     1921 tatcttctat atcttcaggc gcgcggtctg gcagtaaaaa ctatccagca acatttgggc
     1981 cagctaaaca tgcttcatcg tcggtccggg ctgccacgac caagtgacag caatgctgtt
     2041 tcactggtta tgcggcggat ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag
     2101 gctctagcgt tcgaacgcac tgatttcgac caggttcgtt cactcatgga aaatagcgat
     2161 cgctgccagg atatacgtaa tctggcattt ctggggattg cttataacac cctgttacgt
     2221 atagccgaaa ttgccaggat cagggttaaa gatatctcac gtactgacgg tgggagaatg
     2281 ttaatccata ttggcagaac gaaaacgctg gttagcaccg caggtgtaga gaaggcactt
     2341 agcctggggg taactaaact ggtcgagcga tggatttccg tctctggtgt agctgatgat
     2401 ccgaataact acctgttttg ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc
     2461 agccagctat caactcgcgc cctggaaggg atttttgaag caactcatcg attgatttac
     2521 ggcgctaagg atgactctgg tcagagatac ctggcctggt ctggacacag tgcccgtgtc
     2581 ggagccgcgc gagatatggc ccgcgctgga gtttcaatac cggagatcat gcaagctggt
     2641 ggctggacca atgtaaatat tgtcatgaac tatatccgta acctggatag tgaaacaggg
     2701 gcaatggtgc gcctgctgga agatggcgat ggaagcggag agggcagggg aagtcttcta
     2761 acatgcgggg acgtggagga aaatcccggc cccatggtga gcaagggcga ggagctgttc
     2821 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2881 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     2941 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg
     3001 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     3061 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     3121 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3181 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     3241 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     3301 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     3361 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     3421 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3481 atcactctcg gcatggacga gctgtacaag taaacccagc tttcttgtac aaagtgggaa
     3541 ttccgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3601 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3661 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3721 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3781 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3841 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3901 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     3961 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4021 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4081 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     4141 ggaattccta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     4201 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4261 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4321 ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag
     4381 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4441 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     4501 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     4561 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     4621 attaagcgcg gcgggggtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     4681 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4741 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4801 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4861 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4921 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4981 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5041 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     5101 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     5161 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     5221 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     5281 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5341 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5401 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5461 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5521 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5581 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5641 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5701 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5761 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5821 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5881 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5941 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6001 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6061 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6121 ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc
     6181 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6241 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6301 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6361 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6421 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6481 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6541 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6601 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6661 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6721 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6781 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6841 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6901 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6961 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7021 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7081 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
