LOCUS       Exported                6639 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 6639)
  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..6639
                     /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 (oML2-59)
                     /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..2435
                     /codon_start=1
                     /label=VAMP2_DN_mus
                     /note="color: #ffff00"
                     /translation="MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVDEVVDI
                     MRVNVDKVLERDQKLSELDDRADALQAGASQFETSAAKLKRKYWWKNLK"
     primer_bind     complement(2408..2452)
                     /label=oML-Vamp2DN-AgeI-rev
                     /note="color: black; sequence: 
                     cttccgccgctACCGGTCTTGAGGTTTTTCCACCAGTATTTGCGC; added: 
                     2021-04-08"
     primer_bind     2436..2478
                     /label=oML-AgeI-P2AT2A-for
                     /note="color: black; sequence: 
                     ACCGGTagcggcggaagcgctactaacttcagcctgctgaagc; added: 
                     2021-04-08"
     CDS             2454..2510
                     /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 2507"
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(2487..2513)
                     /label=oML-P2AT2A-BamHI-rev
                     /note="color: black; sequence: 
                     catggatccaggtccaggattctcctcgacgtc; added: 2021-04-08"
     primer_bind     complement(2487..2511)
                     /label=oML2-56
                     /note="color: black; sequence: 
                     CATGCTAGCAGGTCCAGGATTCTCCTCGACGTC; added: 2022-07-18"
     CDS             2520..2573
                     /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 2570"
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(2550..2582)
                     /label=oML2-56
                     /note="color: black; sequence: 
                     CATGCTAGCAGGTCCAGGATTCTCCTCGACGTC; added: 2022-07-18"
     primer_bind     complement(2550..2574)
                     /label=oML-P2AT2A-BamHI-rev
                     /note="color: black; sequence: 
                     catggatccaggtccaggattctcctcgacgtc; added: 2021-04-08"
     primer_bind     2565..2601
                     /label=oML-BamHI-dClover-caax-for
                     /note="color: black; sequence: 
                     cctggacctggatccatggtgagcaagggcgaggagc; added: 2021-04-08"
     primer_bind     2565..2601
                     /label=oML2-57
                     /note="color: black; sequence: 
                     CCTGGACCTGCTAGCATGGTGAGCAAGGGCGAGGAGC; added: 2022-07-18"
     CDS             2580..3296
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 2580 .. 2582 / #05fd14
                        2: 2583 .. 2585 / #05fd14 / 1a
                        3: 2586 .. 3296 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2580..2583
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     misc_feature    3297..3371
                     /label=linker-farnesyl tag
                     /note="color: #ffcc00"
     primer_bind     complement(3352..3388)
                     /label=oML-dClover-caax-XhoI-rev
                     /note="color: black; sequence: 
                     cccgtagatctctCGAGTCAGGAGAGCACACACTTGCAGC; added: 
                     2021-04-08"
     polyA_signal    3386..3862
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     misc_feature    complement(3446..3469)
                     /label=C-CMV-24 seq primer
                     /note="Sequetech"
                     /note="color: #a6acb3; direction: LEFT"
     primer_bind     complement(3872..3894)
                     /label=oML2-60
                     /note="color: black; sequence: CTCGGTCCGCACGTGGTTACCTA; 
                     added: 2022-07-18"
     repeat_region   3902..4042
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      4117..4572
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4854..4958
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4959..5819
                     /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: 4959 .. 5027 / #ccffcc / signal sequence
                        2: 5028 .. 5819 / #ccffcc
                     Cleavage site after base 5027"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5990..6578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(6106..6120)
                     /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 accatgtcgg
     2161 ctaccgctgc caccgtcccg cctgccgccc cggccggcga gggtggcccc cctgcacctc
     2221 ctccaaacct tactagtaac aggagactgc agcagaccca ggcccaggtg gatgaggtgg
     2281 tggacatcat gagggtgaat gtggacaagg tcctggagcg ggaccagaag ttgtcggagc
     2341 tggatgaccg tgcagatgcc ctccaggcag gggcctccca gtttgaaaca agtgcagcca
     2401 agctcaagcg caaatactgg tggaaaaacc tcaagaccgg tagcggcgga agcgctacta
     2461 acttcagcct gctgaagcag gctggagacg tggaggagaa ccctggacct ggaagcggag
     2521 agggcagagg gagtctgcta acatgcggtg acgtcgagga gaatcctgga cctgctagca
     2581 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2641 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2701 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2761 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2821 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2881 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2941 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3001 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3061 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3121 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3181 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3241 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg
     3301 gactcagatc taagctgaac cctcctgatg agagtggccc cggctgcatg agctgcaagt
     3361 gtgtgctctc ctgataaaga tctacgggtg gcatccctgt gacccctccc cagtgcctct
     3421 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     3481 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     3541 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     3601 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     3661 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     3721 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     3781 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     3841 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg
     3901 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3961 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4021 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4081 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     4141 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4201 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4261 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4321 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4381 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4441 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     4501 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4561 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4621 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4681 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4741 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4801 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4861 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4921 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4981 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5041 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5101 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5161 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5221 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5281 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5341 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5401 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5461 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5521 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5581 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5641 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5701 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5761 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5821 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5881 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5941 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6001 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6061 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6121 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6181 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6241 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6301 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6361 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6421 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6481 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6541 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6601 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
