LOCUS       pAAV_hSyn_CaBLAM        6388 bp    DNA     circular SYN 04-MAR-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6388)
  AUTHORS   Hochgeschwender
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 2025 from SnapGene 8.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6388
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   5..145
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     promoter        190..637
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     419..441
                     /label=D134
                     /note="color: black; sequence: ACTGAAGGCGCGCTGACGTCACT; 
                     added: 2022-10-18"
     primer_bind     714..730
                     /label=C883
                     /note="color: black; sequence: 
                     gagactctGGATccATGgtgagcaagggc; added: 2024-03-04"
     CDS             716..1393
                     /codon_start=1
                     /label=mNG_d10c
                     /note="color: #00ff00"
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFT"
     regulatory      971..980
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     primer_bind     1178..1197
                     /label=C644
                     /note="color: black; sequence: taccccaacgacaaaaccat; added:
                     2022-10-18"
     primer_bind     complement(1178..1197)
                     /label=C646
                     /note="color: black; sequence: atggttttgtcgttggggta; added:
                     2022-10-18"
     CDS             1394..1399
                     /codon_start=1
                     /label=GF_linker
                     /note="color: #808080"
                     /translation="GF"
     CDS             1400..1858
                     /codon_start=1
                     /label=ekl9h aa 6-156
                     /note="color: #00ffff"
                     /translation="EDFVGDWEQTAAYNLDQVLEQGGVSSVLQTLAVSVTPIQRIVRSG
                     ENGLKIDIHVIIPYEGLSADQMAHIEEVFKVVYPVDDHHFKVIMEYGTLVIDGVTPNML
                     NYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLITPEGSMLFRVTIN"
     primer_bind     1406..1424
                     /label=C608
                     /note="color: black; sequence: TTTCGTTGGGGACTGGGAAC; added:
                     2024-03-04"
     primer_bind     complement(1463..1486)
                     /label=C884
                     /note="color: black; sequence: 
                     agagtctcCTGCAGAACAGAAGAAACACCACC; added: 2024-03-04"
     primer_bind     1480..1497
                     /label=C885
                     /note="color: black; sequence: gagactctCTGCAGACCCTGGCTGT; 
                     added: 2024-03-04"
     primer_bind     1837..1856
                     /label=C650
                     /note="color: black; sequence: GCTGTTCCGTGTTACCATCA; added:
                     2022-10-18"
     CDS             1859..1870
                     /codon_start=1
                     /label=SASS linker
                     /note="color: #808080"
                     /translation="SASS"
     CDS             1871..1936
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /note="color: #ff6600"
                     /translation="DSSRRKWNKTGHAVRAIGRLSS"
     CDS             1937..1981
                     /codon_start=1
                     /label=5xGGSLinker
                     /note="color: #808080"
                     /translation="GGSGGSGGSGGSGGS"
     CDS             1982..2431
                     /codon_start=1
                     /label=Calmodulin
                     /note="color: #ff6600"
                     /translation="NLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRSLGQNP
                     TEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGIGYISA
                     AELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             2432..2440
                     /codon_start=1
                     /label=SPA linker
                     /note="color: #808080"
                     /translation="SPA"
     CDS             2441..2476
                     /codon_start=1
                     /product="sequence from eBlocks_11-15-2020 well D12 
                     APD3A6_LLEEISN"
                     /label=LLEEISN peptide
                     /note="color: #00ffff"
                     /translation="VTGYRLLEEISN"
     primer_bind     complement(2452..2476)
                     /label=C886
                     /note="color: black; sequence: 
                     agagtctcctcgagttattctaaGTTAGAGATTTCTTCGAGCAGACGG; added: 
                     2024-03-04"
     CDS             2510..2527
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     misc_feature    2558..3146
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     primer_bind     complement(2625..2645)
                     /label=D527
                     /note="color: black; sequence: AAGCAGCGTATCCACATAGCG; 
                     added: 2022-10-18"
     polyA_signal    3178..3654
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   3694..3834
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      3909..4364
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4646..4750
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(4751..4819,4820..5611)
                     /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: 4751 .. 4819 / #ccffcc / signal sequence
                        2: 4820 .. 5611 / #ccffcc
                     Cleavage site after base 4819"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5782..6370
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     6250..6269
                     /label=C398
                     /note="color: black; sequence: GCGCACGAGGGAGCTTCCAG; added:
                     2024-03-04"
     primer_bind     6320..6339
                     /label=C397
                     /note="color: black; sequence: CTTGAGCGTCGATTTTTGTG; added:
                     2024-03-04"
     primer_bind     6366..6384
                     /label=C105
                     /note="color: black; sequence: ggaaaaacgccagcaacgc; added: 
                     2024-03-04"
ORIGIN
        1 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
       61 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      121 gccaactcca tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc
      181 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct
      241 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc
      301 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc
      361 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac
      421 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg
      481 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg
      541 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt
      601 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc ggatcctcta
      661 gagtcgagct caccggtggc gcgccaatat tatgctagcg ctaccggtcg ccaccatggt
      721 gagcaagggc gaggaggata acatggcctc tctcccagcg acacatgagt tacacatctt
      781 tggctccatc aacggtgtgg actttgacat ggtgggtcag ggcaccggca atccaaatga
      841 tggttatgag gagttaaacc tgaagtccac caagggtgac ctccagttct ccccctggat
      901 tctggtccct catatcgggt atggcttcca tcagtacctg ccctaccctg acgggatgtc
      961 gcctttccag gccgccatgg tagatggctc cggataccaa gtccatcgca caatgcagtt
     1021 tgaagatggt gcctccctta ctgttaacta ccgctacacc tacgagggaa gccacatcaa
     1081 aggagaggcc caggtgaagg ggactggttt ccctgctgac ggtcctgtga tgaccaactc
     1141 gctgaccgct gcggactggt gcaggtcgaa gaagacttac cccaacgaca aaaccatcat
     1201 cagtaccttt aagtggagtt acaccactgg aaatggcaag cgctaccgga gcactgcgcg
     1261 gaccacctac acctttgcca agccaatggc ggctaactat ctgaagaacc agccgatgta
     1321 cgtgttccgt aagacggagc tcaagcactc caagaccgag ctcaacttca aggagtggca
     1381 aaaggccttt accggatttg aagacttcgt tggtgactgg gaacagaccg ctgcttacaa
     1441 cctggaccag gttctggaac agggtggtgt ttcttctgtt ctgcagaccc tggctgtttc
     1501 tgttacccct atccagcgta tcgttcgttc tggtgaaaac ggtctgaaga tcgacatcca
     1561 tgttatcatc ccttacgaag gtctgtctgc tgaccagatg gctcatatcg aagaagtttt
     1621 caaggttgtt taccctgttg acgaccatca tttcaaggtt atcatggaat acggtaccct
     1681 ggttatcgac ggtgttaccc ctaacatgct gaactacttc ggtcgtcctt acgaaggtat
     1741 cgctgttttc gacggtaaga agatcaccgt taccggtacc ctgtggaacg gtaacaagat
     1801 catcgacgaa cgtctgatca cccctgaggg ttctatgctg ttccgtgtta ccatcaactc
     1861 tgcttcttct gactcttcta ggcgtaagtg gaacaagacc ggtcatgctg ttcgtgctat
     1921 cggtcgtctg tcttctggtg gatccggtgg tagtggtgga tccggaggtt ctggtggatc
     1981 caacctgcct gaccagctga ccgaagaaca gatcgctgaa ttcaaggaag agttctctct
     2041 gttcgataaa gatggagacg gtaccatcac caccaaggaa ctgggtaccg tgatgcgttc
     2101 tctgggtcag aaccctaccg aagctgaact gcaagacatg atcaacgaag ttgacgctga
     2161 cggtgacggt accatcgact tccctgaatt cctgaccatg atggctcgta agatgaagta
     2221 cagggacacc gaagaggaaa tccgtgaagc tttcggcgtt ttcgacaagg acggtatcgg
     2281 ttacatctct gctgctgaac tgcgtcatgt tatgaccaac ctgggtgaaa agctgaccga
     2341 cgaagaagtt gacgaaatga tccgtgaagc tgacatcgac ggtgacggtc aggttaacta
     2401 cgaagaattc gttcagatga tgaccgctaa gtctcctgct gttaccggtt accgtctgct
     2461 cgaagaaatc tctaacctcg agtaacatat gcacgcgtca attgccggtc atcatcacca
     2521 tcaccattga gtttaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     2581 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2641 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2701 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2761 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2821 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2881 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2941 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     3001 gcgcgggacg tccttctgct acgtcccttc ngccctcaat ccagcgganc ttccttcccg
     3061 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agangantcg
     3121 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg
     3181 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3241 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     3301 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     3361 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     3421 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     3481 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     3541 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     3601 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     3661 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     3721 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3781 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     3841 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     3901 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3961 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4021 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4081 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4141 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4201 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt
     4261 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4321 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4381 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4441 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4501 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4561 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4621 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4681 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4741 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4801 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4861 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4921 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4981 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5041 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5101 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5161 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5221 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5281 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5341 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5401 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5461 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5521 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5581 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5641 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5701 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5761 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5821 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5881 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     5941 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6001 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6061 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6121 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6181 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6241 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6301 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6361 cctatggaaa aacgccagca acgcggcc
//
