LOCUS       Exported                4707 bp ds-DNA     circular UNA 04-DEC-2025
DEFINITION  natural linear DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Dec 4, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Template 155A digested with BamHI, NheI 
            f1:gaagcttGCCACCatgCACCTGAGAATCCACGCGAGACGG 
            f2:gtcgtgcctgagagcgcagtcgagaagaagcttGCCACCatgCACCTGAGAATC 
            r1:GTATGGCTCCGGCcAGCTTGCGCCGCGC
            f2:GCGCGGCGCAAGCTgGCCGGAGCCATAC
            r2:CTTATCGTCGTCATCCTTGTAGTCggatc
FEATURES             Location/Qualifiers
     source          1..4707
                     /organism="unspecified"
                     /mol_type="genomic DNA"
                     /note="color: #ffffff"
     source          652..1692
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /note="/locus_tag=source:synthetic DNA construct"
                     /note="color: #ffffff"
     misc_feature    1..130
                     /locus_tag="ITR"
                     /label=ITR
                     /note="color: #a6acb3"
     misc_feature    151..628
                     /locus_tag="synapsin promoter"
                     /label=synapsin promoter
                     /note="color: #a6acb3"
     promoter        181..628
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    637..642
                     /locus_tag="HindIII"
                     /label=HindIII
                     /note="color: #a6acb3"
     regulatory      643..652
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     misc_feature    649..651
                     /locus_tag="Ig k-chain signal sequence"
                     /label=Ig k-chain signal sequence
                     /note="color: #a6acb3"
     misc_feature    652..693
                     /locus_tag="V5 tag"
                     /label=V5 tag
                     /note="color: #a6acb3"
     CDS             652..693
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /note="color: #cc99b2"
                     /translation="GKPIPNPLLGLDST"
     misc_feature    694..699
                     /locus_tag="NheI"
                     /label=NheI
                     /note="color: #a6acb3"
     CDS             700..1656
                     /codon_start=1
                     /locus_tag="BirA (humanized)"
                     /label=BirA (humanized)
                     /note="color: #993366"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     misc_feature    889..891
                     /locus_tag="Q65P"
                     /label=Q65P
                     /note="color: #a6acb3"
     misc_feature    955..957
                     /locus_tag="I87V"
                     /label=I87V
                     /note="color: #a6acb3"
     misc_feature    1039..1041
                     /locus_tag="G115G"
                     /label=G115G
                     /note="color: #a6acb3"
     misc_feature    1048..1050
                     /locus_tag="R118S"
                     /label=R118S
                     /note="color: #a6acb3"
     misc_feature    1114..1116
                     /locus_tag="E141K"
                     /label=E141K
                     /note="color: #a6acb3"
     misc_difference 1117..1119
                     /locus_tag="Q142R"
                     /label=Q142R
                     /note="color: #a6acb3"
     misc_feature    1141..1143
                     /locus_tag="S150G"
                     /label=S150G
                     /note="color: #a6acb3"
     misc_feature    1144..1146
                     /locus_tag="L151P"
                     /label=L151P
                     /note="color: #a6acb3"
     misc_feature    1171..1173
                     /locus_tag="V160A"
                     /label=V160A
                     /note="color: #a6acb3"
     misc_feature    1267..1269
                     /locus_tag="T192A"
                     /label=T192A
                     /note="color: #a6acb3"
     misc_difference 1273..1275
                     /locus_tag="K194I"
                     /label=K194I
                     /note="color: #a6acb3"
     misc_feature    1318..1320
                     /locus_tag="M209V"
                     /label=M209V
                     /note="color: #a6acb3"
     misc_feature    1414..1416
                     /locus_tag="M241T"
                     /label=M241T
                     /note="color: #a6acb3"
     misc_feature    1480..1482
                     /locus_tag="S263P"
                     /label=S263P
                     /note="color: #a6acb3"
     misc_structure  1564..1566
                     /locus_tag="G291G"
                     /label=G291G
                     /note="color: #a6acb3"
     misc_feature    1606..1608
                     /locus_tag="I305V"
                     /label=I305V
                     /note="color: #a6acb3"
     misc_feature    join(1657..1667,1669..1692)
                     /locus_tag="NES"
                     /label=NES
                     /note="This feature has 2 segments:
                        1: 1657 .. 1667 / #a6acb3
                        2: 1669 .. 1692 / #a6acb3"
     CDS             1663..1689
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /note="color: #993366"
                     /translation="LPPLERLTL"
     misc_feature    complement(1693..1695)
                     /locus_tag="double stop"
                     /label=double stop
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    1699..1704
                     /locus_tag="EcoRI"
                     /label=EcoRI
                     /note="color: #a6acb3"
     misc_feature    1711..1942
                     /locus_tag="PolyA"
                     /label=PolyA
                     /note="color: #a6acb3"
     polyA_signal    1735..1942
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     misc_feature    1971..2111
                     /locus_tag="ITR(1)"
                     /label=ITR
                     /label=ITR(1)
                     /note="color: #a6acb3"
     repeat_region   1971..2111
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      2185..2640
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        2922..3026
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3027..3887
                     /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: 3027 .. 3095 / #ccffcc / signal sequence
                        2: 3096 .. 3887 / #ccffcc
                     Cleavage site after base 3095"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4058..4646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg
      181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      601 aggagtcgtg tcgtgcctga gagcgcagtc gagaagaagc ttgccaccat gggcaagccc
      661 atccccaacc ccctgctggg cctggacagc accgctagca aagacaatac tgtgcctctg
      721 aagctgatcg ctctcctggc taatggcgag ttccatagtg gcgaacagct gggagaaacc
      781 ctgggcatgt ccagggccgc tatcaacaag cacattcaga ctctgcgcga ctggggcgtg
      841 gacgtgttca ccgtgcccgg aaagggctac tctctgcccg agcctatccc gctgctgaac
      901 gctaaacaga ttctgggaca gctggacggc gggagcgtgg cagtcctgcc tgtggtcgac
      961 tccaccaatc agtacctgct ggatcgaatc ggcgagctga agagtgggga tgcttgcatt
     1021 gcagaatatc agcaggcagg gagaggaagc agagggagga aatggttctc tccttttgga
     1081 gctaacctgt acctgagtat gttttggcgc ctgaagcggg gaccagcagc aatcggcctg
     1141 ggcccggtca tcggaattgt catggcagaa gcgctgcgaa agctgggagc agacaaggtg
     1201 cgagtcaaat ggcccaatga cctgtatctg caggatagaa agctggcagg catcctggtg
     1261 gagctggccg gaataacagg cgatgctgca cagatcgtca ttggcgccgg gattaacgtg
     1321 gctatgaggc gcgtggagga aagcgtggtc aatcagggct ggatcacact gcaggaagca
     1381 gggattaacc tggacaggaa tactctggcc gctacgctga tccgagagct gcgggcagcc
     1441 ctggaactgt tcgagcagga aggcctggct ccatatctgc cacggtggga gaagctggat
     1501 aacttcatca atagacccgt gaagctgatc attggggaca aagagatttt cgggattagc
     1561 cgggggattg ataaacaggg agccctgctg ctggaacagg acggagttat caaaccctgg
     1621 atgggcggag aaatcagtct gcggtctgcc gaaaagctgc agctgcctcc cctggagcgc
     1681 ctgaccctgg actaatagga attcctcgag ctagagctcg ctgatcagcc tcgactgtgc
     1741 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1801 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1861 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     1921 acaatagcag gcatgctggg gaggtaacca cgtgcggacc gagcggccgc aggaacccct
     1981 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     2041 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2101 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtattcacac
     2161 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2221 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2281 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2341 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2401 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2461 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2521 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2581 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     2641 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2701 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2761 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2821 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     2881 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2941 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3001 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3061 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3121 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3181 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3241 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3301 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3361 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3421 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3481 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3541 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3601 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3661 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3721 aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg ccagatggta
     3781 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3841 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3901 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3961 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4021 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4081 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4141 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4201 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4261 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4321 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4381 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4441 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4501 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4561 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4621 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4681 ccttttgctg gccttttgct cacatgt
//
