LOCUS       sequence-479484-        7104 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  5' AAVLINK plasmid for RALGAPB expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  AUTHORS   Zhonghua Lu Lab
  TITLE     Zhonghua Lu AAV Plasmids 2025
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7104)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAVLINK-EF1alpha-5' RALGAPB
FEATURES             Location/Qualifiers
     source          1..7104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..386
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             399..667
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             704..730
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     RBS             1372..1380
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     repeat_region   4367..4507
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4367..4496
                     /label=AAV2 ITR
     rep_origin      4582..5037
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4669..4688)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4879..4900
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5054..5073)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5173..5195
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5233..5251)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5319..5423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5424..5492,5493..6284)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5642..5661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6455..7043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6944..6963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtggatct gcgatcgctc cggtgcccgt cagtgggcag
      181 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
      241 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      301 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      361 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
      421 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
      481 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
      541 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
      601 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
      661 acagatccaa gctgtgaccg gcgcctaccg tacggccacc atgtaccctt atgacgtacc
      721 tgactatgct ggatcttact ctgagtggag gtcactgcat ttggtgattc agaatgatca
      781 aggccatacc agtgtgctgc acagctatcc agagagcgtt ggacgagagg tggcaaatgc
      841 tgtagtccgt cctcttgggc aggtgttagg taccccttca gtggctggta gtgagaattt
      901 gttaaaaact gacaaagaag taaaatggac catggaagta atttgctatg gactgaccct
      961 tccattggat ggagagactg taaaatattg cgttgatgta tatacagact ggattatggc
     1021 tttagtgttg ccaaaagatt ctattccatt gccagttatt aaagagccta atcaatatgt
     1081 tcaaactata ctaaaacacc tacagaatct ttttgtacca agacaggaac agggttccag
     1141 tcagattcga ctatgcttac aggtcctgag agccattcag aaactggccc gtgagtcatc
     1201 tctcatggcc cgagaaactt gggaagtctt actgttgttt cttctgcaga ttaacgacat
     1261 acttctggcc ccaccaactg ttcaaggtgg cattgctgag aatctagcag agaagttgat
     1321 tggtgttctc tttgaggtgt ggttactagc ttgtactcgg tgcttcccaa cacctcctta
     1381 ttggaaaaca gccaaggaga tggtggctaa ctggaggcat cacccagcag tggtggagca
     1441 gtggagcaag gtcatttgtg cactcacttc cagattgcta cgctttacat atggtccttc
     1501 atttcctgca tttaaagttc ccgatgaaga tgccagtctg atccctccag aaatggataa
     1561 tgagtgtgtt gcacagacat ggtttcgctt tttacacatg ttaagtaatc ctgtggattt
     1621 gagtaaccca gctattataa gctctactcc caaatttcag gaacagttct tgaatgtgag
     1681 cggaatgccg caagaattga atcagtatcc ctgccttaaa catctgcctc aaatattttt
     1741 tcgtgccatg cgtggaatca gctgtctggt ggatgcattc ttaggtattt ctagaccccg
     1801 atcagacagt gctcccccaa cacccgtgaa tagattaagt atgcctcaaa gtgctgctgt
     1861 cagtaccacc cccccacata accggaggca ccgggctgtt actgtgaata aggccaccat
     1921 gaagacaagc acagttagta ctgctcatgc ctctaaagtt cagcaccaga cgtcctccac
     1981 ctctcctctg tcaagtccaa atcagactag ttcagaaccc cggccactgc ctgcccctcg
     2041 gagaccaaag gttaacagca tcttgaatct ctttggatca tggttatttg atgcagcatt
     2101 tgttcactgt aaacttcata atgggataaa cagagacagc agcatgactg ccattacaac
     2161 acaagctagc atggagtttc gacggaaagg gtcacaaatg tccacagaca ccatggtttc
     2221 caatcctatg tttgatgcaa gtgaatttcc tgataactat gaagcaggaa gagctgaggc
     2281 ttgtgggaca ctgtgtagga ttttttgtag caagaagact ggagaagaga ttctgccagc
     2341 ttatttatcc agattttaca tgcttttaat tcaaggtttg cagataaatg attatgtgtg
     2401 ccatcctgtc ttggccagcg ttattctaaa ctctcctcct ttgttctgct gtgacttgaa
     2461 agggattgat gttgtggttc cttactttat ttcagctctt gaaaccattt tgcctgacag
     2521 agaactctca aaattcaaaa gctatgtaaa tccaacagaa ttgcgaagat cctccattaa
     2581 tatcctgctt tctttgttgc ccctccctca tcattttggc acagtcaaat ctgaggtggt
     2641 cctggaagga aagtttagta acgatgacag ctcttcttat gataaaccaa taacttttct
     2701 gtccctgaag ttgagacttg tgaatatatt aataggtgcc ttgcaaactg aaacggaccc
     2761 caacaacacc caaatgatat taggggcaat gttaaatatt gttcaagatt cagcactttt
     2821 ggaagccatt ggttgccaga tggagatggg tggtggagaa aataacctga agagtcatag
     2881 tcgcaccaat agtggtatta gttcagcaag tggtggaagc acggagccca cgactcccga
     2941 tagtgagaga cctgctcaag ctctcttaag agattatgct cttaatacag attcagctgc
     3001 tgggctcctg attcgcagca ttcatctcgt cacccaaaga ctcaactccc agtggcgcca
     3061 agacatgagc atatcactgg cagctctaga gctcctctct ggccttgcaa aggtaaaagt
     3121 gatggttgac tcaggagacc ggaagcgagc catcagttct gtgtgcacct acattgttta
     3181 tcagtgtagt cggccagctc ctttacactc cagggatctg cactccatga tagtggcagc
     3241 ttttcagtgt ctctgtgtct ggctgacaga gcaccctgat atgcttgatg aaaaggactg
     3301 ccttaaggaa gtactggaga ttgtggaact gggtatctca ggaagtaagt ccaagaacaa
     3361 tgagcaagag gtcaagtaca aaggagataa ggagccaaac cctgcatcta tgagggtaaa
     3421 ggatgctgct gaagccaccc taacatgcat tatgcagttg ctcggcgcat ttccttcacc
     3481 tagtggtcct gcctctcctt gtagtcttgt gaatgagacc actttgatta aatactccag
     3541 gctgccaacc ataaacaagc atagtttccg gtactttgtc ttggataaca gtgtcatcct
     3601 ggcaatgctg gaacaacctc ttggaaatga gcagaatgat tttttcccct ctgtcactgt
     3661 gctggtccgg ggaatgtctg gaagacttgc ttgggcacaa cagctttgtc ttttacccag
     3721 aggagcaaaa gcaaatcaga agctttttgt acctgaacct cgcccagttc ctaaaaatga
     3781 cgttggattt aaatattctg tgaaacatcg gccatttcct gaagaggtgg acaagattcc
     3841 ttttgtgaaa gcagatctca gcattccaga tttgcatgaa atagtcactg aagaattaga
     3901 agagagacac gaaaaattaa ggagtggcat ggcccagcag attgcttatg aaatacacct
     3961 tgagcaacag agtgaggagg aattgcagaa gagaagtttt cctgacccag ttacggattg
     4021 caagcccccg cctcctgccc aggaattcca aacagcccgc ctttttctct cacactttgg
     4081 atttttgtcc ttagaagcac tgaaggaacc tgcaaatagt cgtctacctc ctcaccttat
     4141 tgcacttgat tccacgatac ctggattttt tgatgacatt gggtatctgg atctcttgcc
     4201 atgtcgtcct tttgacacag tttttatttt ctatatgaag ccaggtaagt atcaaggtta
     4261 caagacaggt ttaaggagac caataagcta ttattcgtat aggatacttt atacgaagtt
     4321 attaagttat ggagaccggt gtggcacgtg cggaccgagc ggccgcagga acccctagtg
     4381 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4441 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4501 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4561 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4621 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     4681 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4741 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4801 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4861 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     4921 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     4981 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     5041 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     5101 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5161 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5221 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5281 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5341 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5401 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5461 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5521 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5581 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5641 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5701 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5761 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5821 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5881 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5941 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6001 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6061 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6121 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6181 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6241 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6301 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6361 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6421 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6481 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6541 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6601 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6661 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6721 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6781 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6841 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6901 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6961 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7021 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7081 tttgctggcc ttttgctcac atgt
//
