LOCUS       Exported                5697 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 5697)
  AUTHORS   Brad Zuchero
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Aug 16, 2022 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5697
                     /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"
     enhancer        221..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        525..727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     720..738
                     /label=Sequetech""VectorPrimer3""
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    748..765
                     /label=pTRE5 seq primer
                     /note="Sequetech"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    871..1254
                     /label=CMV / beta-globin hybrid intron (Not necessary!)
                     /note="color: #a6acb3"
     misc_feature    1255..1307
                     /label=globin exon 3 (partial) Not necessary!
                     /note="color: #a6acb3"
     regulatory      1332..1337
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             1338..1685
                     /codon_start=1
                     /label=Vamp2
                     /note="color: #ffff00"
                     /translation="MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVDEVVDI
                     MRVNVDKVLERDQKLSELDDRADALQAGASQFETSAAKLKRKYWWKNLKMMIILGVICA
                     IILIIIIVYFST"
     misc_feature    1686..1715
                     /label=GS linker
                     /note="color: #a6acb3"
     CDS             1716..2426
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /note="color: #05fd14"
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKANF
                     KIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELYK"
     polyA_signal    2444..2920
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     misc_feature    complement(2457..2474)
                     /label=PCMV5R seq primer
                     /note="Sequetech"
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    complement(2504..2527)
                     /label=C-CMV-24 seq primer
                     /note="Sequetech"
                     /note="color: #a6acb3; direction: LEFT"
     repeat_region   2960..3100
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      3175..3630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3912..4016
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4017..4877
                     /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: 4017 .. 4085 / #ccffcc / signal sequence
                        2: 4086 .. 4877 / #ccffcc
                     Cleavage site after base 4085"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5048..5636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtggagc tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgtcaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttgcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggattc gaatcccggc cgggaacggt
      841 gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata
      901 ggcccacaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac
      961 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc
     1021 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata
     1081 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta
     1141 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt
     1201 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg
     1261 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagaattggg attcgaacat
     1321 cgattgaatt cgccaccatg tcggctaccg ctgccaccgt cccgcctgcc gccccggccg
     1381 gcgagggtgg cccccctgca cctcctccaa accttactag taacaggaga ctgcagcaga
     1441 cccaggccca ggtggatgag gtggtggaca tcatgagggt gaatgtggac aaggtcctgg
     1501 agcgggacca gaagttgtcg gagctggatg accgtgcaga tgccctccag gcaggggcct
     1561 cccagtttga aacaagtgca gccaagctca agcgcaaata ctggtggaaa aacctcaaga
     1621 tgatgatcat cttgggagtg atctgcgcca tcatcctcat catcatcatc gtttacttca
     1681 gcactagcgg cggaagcggc gggaccggtg gaagtaaagg agaagaactt ttcactggag
     1741 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     1801 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     1861 gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt gttcaatgct
     1921 tttcaagata cccagatcat atgaaacggc atgacttttt caagagtgcc atgcccgaag
     1981 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg
     2041 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     2101 aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataacgat caccaggtgt
     2161 acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt agacacaaca
     2221 ttgaagatgg aggcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     2281 ggcccgtcct tttaccagac aaccattacc tgtttacaac ttctactctt tcgaaagatc
     2341 ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac
     2401 atggcatgga tgaactatac aaataatgac tcgagagatc tacgggtggc atccctgtga
     2461 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     2521 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     2581 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     2641 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     2701 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     2761 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     2821 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     2881 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     2941 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3001 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3061 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     3121 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3181 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3241 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3301 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3361 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3421 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3481 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     3541 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3601 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3661 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3721 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3781 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3841 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3901 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3961 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4021 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4081 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4141 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4201 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4261 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4321 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4381 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4441 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4501 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4561 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4621 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4681 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4741 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4801 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4861 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4921 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4981 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5041 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5101 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5161 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     5221 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5281 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5341 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5401 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5461 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5521 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5581 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5641 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
