LOCUS       Exported                6600 bp ds-DNA     circular SYN 16-AUG-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6600)
  AUTHORS   JBZ
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Aug 16, 2022 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6600
                     /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"
                     /note="color: #ffe4c4"
     misc_feature    156..2108
                     /label=MBPpromoter
                     /note="Lazzarini"
                     /note="color: #ff99cc"
     primer_bind     2117..2158
                     /label=oML-BamHI-VAMP-for
                     /note="color: black; sequence: 
                     TAATTCTAGGATCCGAACATCGATTGAATTCGCCACCATGTC; added: 
                     2021-04-08"
     primer_bind     2130..2153
                     /label=oML-EcoRI-Vamp-for
                     /note="color: black; sequence: 
                     gattcgaacatcgattgaattCGCCACC; added: 2021-04-08"
     regulatory      2148..2153
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             2154..2396
                     /codon_start=1
                     /label=Vamp3 (rat)-DN
                     /note="color: #993366"
                     /translation="MSTGVPSGSSAATGSNRRLQQTQNQVDEVVDIMRVNVDKVLERDQ
                     KLSELDDRADALQAGASQFETSAAKLKRKYWWKNCK"
     primer_bind     complement(2367..2411)
                     /label=oML-Vamp3DN-AgeI-rev
                     /note="color: black; sequence: 
                     tccgccgctACCGGTCTTGCAGTTCTTCCACCAATACTTTCTCTT; added: 
                     2021-04-08"
     primer_bind     2397..2439
                     /label=oML-AgeI-P2AT2A-for
                     /note="color: black; sequence: 
                     ACCGGTagcggcggaagcgctactaacttcagcctgctgaagc; added: 
                     2021-04-08"
     CDS             2415..2471
                     /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."
                     /note="color: #cc99b2
                     Cleavage site after base 2468"
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(2448..2474)
                     /label=oML-P2AT2A-BamHI-rev
                     /note="color: black; sequence: 
                     catggatccaggtccaggattctcctcgacgtc; added: 2021-04-08"
     CDS             2481..2534
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #cc99b2
                     Cleavage site after base 2531"
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(2511..2535)
                     /label=oML-P2AT2A-BamHI-rev
                     /note="color: black; sequence: 
                     catggatccaggtccaggattctcctcgacgtc; added: 2021-04-08"
     primer_bind     2526..2562
                     /label=oML-BamHI-dClover-caax-for
                     /note="color: black; sequence: 
                     cctggacctggatccatggtgagcaagggcgaggagc; added: 2021-04-08"
     CDS             2541..3257
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 2541 .. 2543 / #05fd14
                        2: 2544 .. 2546 / #05fd14 / 1a
                        3: 2547 .. 3257 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     2541..2562
                     /label=oML-BspEI-mClover
                     /note="color: black; sequence: 
                     CTTCAGCACTTCCGGAATGGTGAGCAAGGGCGAGGAGC; added: 2021-03-12"
     regulatory      2541..2544
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     primer_bind     complement(2586..2607)
                     /label=GFPReverse
                     /note="Sequetech"
                     /note="color: green; sequence: CGTCGCCGTCCAGCTCGACCAG; 
                     added: 2019-03-18"
     misc_feature    3258..3332
                     /label=linker-farnesyl tag
                     /note="color: #ffcc00"
     primer_bind     complement(3313..3349)
                     /label=oML-dClover-caax-XhoI-rev
                     /note="color: black; sequence: 
                     cccgtagatctctCGAGTCAGGAGAGCACACACTTGCAGC; added: 
                     2021-04-08"
     polyA_signal    3347..3823
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     misc_feature    complement(3407..3430)
                     /label=C-CMV-24 seq primer
                     /note="Sequetech"
                     /note="color: #a6acb3; direction: LEFT"
     repeat_region   3863..4003
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      4078..4533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4815..4919
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4920..5780
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 4920 .. 4988 / #ccffcc / signal sequence
                        2: 4989 .. 5780 / #ccffcc
                     Cleavage site after base 4988"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5951..6539
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(6067..6081)
                     /label=oML-AgeI-P2AT2A-for
                     /note="color: black; sequence: 
                     ACCGGTagcggcggaagcgctactaacttcagcctgctgaagc; added: 
                     2021-04-08"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgagct ccttcctgct taggccgtgt
      181 ctgctctgtc agtctttcct ctggctctag tctgtccatc tgcagggata acctcccaag
      241 ggccctcttg ttggctgttc accttccgtc tgtcttcata gcctcctgat gagagagcgc
      301 gcacacacag gctacctgac tgatgtgttg ctgcactgtg gggtccagtg acacagctgt
      361 ggaggtcagg aacccagaat taagctgtct gggagccaca agtttcaaga gtccatgcta
      421 tggctcctac atgcaggtgg tgccagcagc attccttggt gtcctgtggc tgtggttgca
      481 tctctgactc tcctccacct atcacattcc ttctccagta ccaggcctct gtgtctcagt
      541 tttcctctct cacaagatcc ctagcttcag gattgcaccc acctccacta gccatgagcc
      601 cgtcctcact ggatcacgtc tgcaaagact ctatttccag ttaaagtcac agacacgggc
      661 atagagaggg ggataggata tgaaaatatc agttggggga ggggcattca attcttagct
      721 cacattttct tcatccaatt tcccccacta gaatgtaagc tttgagagaa aagggaccag
      781 atcttattcc tcaccgtggc tttaacactt agagaaaatg catcccctct aatcaataag
      841 tcatcgacag tgggtagatg gaggaacggc agtgcgtagt aggatgcgtg caagcatagt
      901 ctcgtgcatg ggtgcataga tcgctgggca ggtggacaag gtgggggtgg ataaagaagt
      961 gggtagatga ttgatgttag gtaaatatca ctgggtggac agatgggtgg taggtggatg
     1021 gatggttaga atagtcagaa gagggatgga ttgataaggt gaacagatga taaatgggtg
     1081 atagactgga agggttgtca aaagaggata agggaagtgt gagctagccg tatttctaag
     1141 gtcagtaata gagttgggag aagaggttaa gttacatcca tttaaacctc acacgaagct
     1201 gagagggaat ggacttgctg ccgttggtga ggaaagcgtt gcatttcccg tgtgcttggt
     1261 tgtgaagtgc tcaggtccca catgaagcag tcaggttact gcggcttaca gaggagccag
     1321 atccaaatgc cccgagtaag cacgtccccg agccagaggc ctccagcgga atccgggaga
     1381 gggattgctc agtgccctgc ttccctggac tgtaagctgc agaaagatgt gggaagtcct
     1441 gttctccact gagaacacta aaagcacctt ttgtcaaacg accgcttcac atctggggct
     1501 tgtgcactgg tggcctttta aaccagagac aacccacaag atacctaacc tgcggggctc
     1561 tctggtacag tgagcaactc aggaaatgct ttggcttgat tgctgtgggc tctcaggcca
     1621 tcgccctctg gagtggttct tttaatgaga acctgaagat tggcccctga gccatgtata
     1681 ccaagcaagc tcaatccagg ttagctccct ctggttgggg caagctaacg tgctccttgg
     1741 gccccgcgcg taactgtgcg ttttatagga gacagctagt tcaagacccc aggaagaaag
     1801 cggctttgtc cccctctagg cctcgtacag gcccacattc atatctcatt gttgttgcag
     1861 gggaggcaga tgcgatccag aacaatggga cctcggctga ggacacggcg gtgacagact
     1921 ccaagcacac agcagaccca aagaataact ggcaaggcgc ccacccagct gacccaggga
     1981 accgccccca cttgatccgc ctcttttccc gagatgcccc gggaagggag gacaacacct
     2041 tcaaagacag gccctcagag tccgacgagc ttcagaccat ccaagaagac cccacagcag
     2101 cttccggaag ctcgaataat tctaggatcc gaacatcgat tgaattcgcc accatgtcta
     2161 caggggtgcc ttcagggtcc agtgctgcca ctggcagtaa tcgaagactc cagcagacac
     2221 agaatcaagt agatgaggtg gtggacatca tgagagtcaa tgtggataag gtgttagaaa
     2281 gagaccagaa gctctcagag ctagacgacc gcgcagatgc actgcaggca ggcgcctcgc
     2341 agtttgaaac aagtgctgcc aagttgaaga gaaagtattg gtggaagaac tgcaagaccg
     2401 gtagcggcgg aagcgctact aacttcagcc tgctgaagca ggctggagac gtggaggaga
     2461 accctggacc tggaagcgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg
     2521 agaatcctgg acctgctagc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2581 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2641 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2701 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     2761 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2821 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2881 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2941 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3001 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3061 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3121 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     3181 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3241 tggacgagct gtacaagtcc ggactcagat ctaagctgaa ccctcctgat gagagtggcc
     3301 ccggctgcat gagctgcaag tgtgtgctct cctgataaag atctacgggt ggcatccctg
     3361 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3421 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3481 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3541 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3601 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3661 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3721 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3781 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     3841 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3901 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3961 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4021 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4081 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4141 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4201 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4261 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4321 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4381 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     4441 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4501 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4561 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4621 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4681 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4741 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4801 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4861 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4921 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4981 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5041 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5101 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5161 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5221 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5281 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5341 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5401 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5461 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5521 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5581 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5641 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5701 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5761 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5821 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5881 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5941 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6001 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6061 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     6121 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6181 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6241 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6301 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6361 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6421 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6481 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6541 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
