LOCUS       Exported                8068 bp DNA     circular SYN 29-SEP-2021
DEFINITION  2 fragments assembled using overlaps.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8068)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 29, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8068
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..186
                     /label=ITR
     misc_feature    193..2734
                     /label=mGAD65(delE1)
     regulatory      2748..2757
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2754..3473
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    3489..4077
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3960..3971)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    4099..4228
                     /label='SV40 polyA
     misc_feature    4251..4436
                     /label=ITR
     rep_origin      complement(4919..5432)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6111..6215
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6216..7076
                     /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      7247..7835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctagactcg aggctttgct tcctgggcac tacagcccca cctgaagcag gtgcgttagg
      241 gaggagttgg gctctgcagc ttgccagatt ttaattctgg cttccagcgg aggaccctgc
      301 accccacggt cctcgcaaga gtgtctggga aggtttattc ggttgctgag tagagacgca
      361 acagacagat ctacctcagc ccctgcacct tggcttgata aggctggaaa aggaagattg
      421 caacgtcgtg gttgtttcca aacccttttt ctatctagcc tttttcagtc tctcaacatt
      481 taagcagcaa aagaactcga gagagaaaac ttgtactaaa tatccttctt tgaccggccc
      541 tctcctctcc ccactacggg gatttctggg tggctcacag tggacaagga ataggtaatc
      601 ccaccaaacc ttgaggtggg ctaaactgga gattttggta tgggtctcct actgtaaaga
      661 gaaagccttc cgggctaacg gtattccttt cgcagtctac tgcctccttt gtgaactaat
      721 gactgtaatt attacctccc agagctcttt tgttatcaac aacccggagc cagagggggg
      781 tgggggatgg gccctttggg aaaagggaaa aaaatttaca aagcaaattg ctgtctacga
      841 agggttggtt ttttaaaaag acaaacccaa ttccagcctg cattcgaagt ggggagtggc
      901 aaaatttttt ttcaaagtat gataagattt taaaaatact cagagctgtg ggattcaatt
      961 gcatttccac agtatcgctt taagttacta tttttgacga agtaaaattc agatgttgaa
     1021 aatctgtatc tcttgttttc atccatttct tccttggaag gtgtgtgtgt gtgtgtgtgt
     1081 gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtttaa gaaccttttt tttttatttc
     1141 cgaccactca tccatggtaa aaagcttcaa ccccagccag ctggcttgaa gagaattatt
     1201 ttgcaggatt tagtatggat ttaaacaaag actcaggctc tccaagtctt gttggcaagg
     1261 ctgggcggaa gcgccctttc caggagctgt tacctctgcc agctgaagcc accaggcctg
     1321 gaggacccta aagacttact aagttgaggc ctagggtcaa ggtgactgcc ttcccttaga
     1381 gacacgcctt gggtagcacc tcagcagtgc caatctcacc gccctccccc tccccacctc
     1441 gggccactca ccggacctca gtgggcatcc tctagctaga tcctccctgg cccccctgga
     1501 ttctggaaac cttgggaaga agataaagct ttgtggagag aatgcacttg gtcatctgca
     1561 gttattcgag gtctccggat ccctaacaaa ataagtgatc ctggtcacca agactaaagg
     1621 tcccagggtt tctcctccgg caacagcttt ggtgactcat ttgaaaaact caaagaagga
     1681 ataaaggact atggagagca gggaagctgc agtccctcgc tcggagagaa ggaaaggaga
     1741 tcgggaacgg tttaaagcag cactttcctg caagtctttc ttgagctctc tgtaagcctg
     1801 gaactatggg tttagtatag gcaagaggaa acggaagcgg agccagaggg ccccaaatct
     1861 tcctcaaggc gccgtccctg catttaagag cctttgggaa aagggaaaca gaaaggattt
     1921 tttcattcct ttattagaaa acacaaacga aactacttct ttctgggctg ggaaattgcc
     1981 actgttgtgt tcctaggcac ctgctgatgg caagaactcc tcttgggaag ggacgaggtc
     2041 agtgggttga gtatccctga aaagatggga gtgccttttc ctgggatttg gaaaaactga
     2101 ttcgcttact ttgagactcc ctgaatcctg tgtatgtgtc agctctcacc cactccaaga
     2161 agtttctgtt cgttttattt atttaatttt tctcagcctg aggtcctcag tgatagattc
     2221 cagcgtggat tttaattgct tcaatcagca gcctttctct tcagccgtca gtcaaaacct
     2281 ggacgatggg tctggggacc caggcatgac tgcacagaaa ccgtacgacc cccaccccca
     2341 ccccccaggc tggctcagct cagcaggcag acattaggct ctgctaagga gatgaaatga
     2401 gttcgttggt gtggaagcgg tccaccttcc tccctccttg gttccttccc caagctttgg
     2461 agctcttcct gattgaggcc aggtggcctg ggctcttgag gtcacagcga cctcagcact
     2521 ggtttggagg aaggcgggcg cgagaaccac ttttgctcgg cagcagccag agacagccct
     2581 caaacaatga gccaatcgat gcacacgtga aattgccctc ggccaccatt ccaatcttgc
     2641 caaggtctcc ctgtccctgg ggtcacctag gccagcgtgc tgagggctgt ggtagcaccc
     2701 tggacagcgc gttccggcag aagcagacct ggaggaattc accggtcgcc accatggtga
     2761 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     2821 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     2881 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2941 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     3001 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     3061 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     3121 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     3181 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     3241 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     3301 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     3361 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     3421 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     3481 gcatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3541 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3601 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3661 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3721 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3781 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3841 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     3901 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3961 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     4021 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     4081 gatgccgtat acggtaccac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4141 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4201 aactcatcaa tgtatcttat catgtctgtc gacccccaac gcgttggagc tctagagcat
     4261 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     4321 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4381 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctggcgt
     4441 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4501 tggaanttcc aacatccaat aaatcataca ggcaaggcaa agaattagca aaattaagca
     4561 ataaagcctc agagcataaa gctaaatcgg ttgtaccaaa aacattatga ccctgtaata
     4621 cttttgcggg agaagccttt atttcaacgc aaggataaaa atttttagaa ccctcatata
     4681 ttttaaatgc aatgcctgag taatgtgtag gtaaagattc aaacgggtga gaaaggccgg
     4741 agacagtcaa atcaccatca atatgatatt caaccgttct agctgataaa ttcatgccgg
     4801 agagggtagc tatttttgag aggtctctac aaaggctatc aggtcattgc ctgagagtct
     4861 ggagcaaaca agagaatcga tgaacggtaa tcgtaaaact agcatgtcaa tcatatgtac
     4921 cccggttgat aatcagaaaa gccccaaaaa caggaagatt gtataagcaa atatttaaat
     4981 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     5041 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     5101 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     5161 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     5221 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     5281 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     5341 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     5401 cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg agcacgtata
     5461 acgtgctttc ctcgttagaa tcagagcggg agctaaacag gaggccgatt aaagggattt
     5521 tagacaggaa cggtacgcca gaatcctgag aagtgttttt ataatcagtg aggccaccga
     5581 gtaaaagagt ctgtccatca cgcaaattaa ccgttgtcgc aatacttctt tgattagtaa
     5641 taacatcact tgcctgagta gaagaactca aactatcggc cttgctggta atatccagaa
     5701 caatattacc gccagccatt gcaacaggaa aaacgctcat ggaaatacct acattttgac
     5761 gctcaatcgt ctggaanttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     5821 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     5881 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     5941 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     6001 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     6061 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6121 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6181 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6241 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6301 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6361 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6421 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6481 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6541 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6601 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6661 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6721 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6781 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6841 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6901 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6961 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7021 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7081 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7141 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7201 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7261 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7321 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7381 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7441 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7501 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7561 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7621 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7681 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7741 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7801 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7861 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     7921 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     7981 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     8041 tccccgcgcg ttggccgatt cattaatg
//
