LOCUS       sequence-414378-        7337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing pAce in a Flp-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7337)
  TITLE     AAV constructs for expressing genetically encoded voltage indicators
            in mice
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7337)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HT152_pAAV_EF1a_fDiO_pAce
FEATURES             Location/Qualifiers
     source          1..7337
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1453..1500
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      2340..2349
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3249..3258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3263..3310)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    3464..4052
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3517..3537)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3935..3946)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4084..4560
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4283..4302)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4600..4740
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4600..4729
                     /label=AAV2 ITR
     rep_origin      4815..5270
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4902..4921)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5112..5133
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5287..5306)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5406..5428
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5466..5484)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5552..5656
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5657..5725,5726..6517)
                     /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(5875..5894)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6688..7276
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7177..7196
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gacggctgtg
     1441 ggcccgtaac aagaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc
     1501 atcaaaatag gaagaccaat gcttcaccat cgacccgaat tgccaagcat caccatcgac
     1561 agaagttcct attccgaagt tcctattctt caaaaggtat aggaacttcg gcgcgcctta
     1621 gaatcgagta gcctcgggga aatctgttgc gcaagagatg aagctatcaa tagatgacat
     1681 actcctcatg tcaatcactc cttcagtcct ggcagggagg ggagccacgg gacttggcat
     1741 actggacatc tcaagctctt cgggaggctt agactgtgga gccatttcct ttgtgttcag
     1801 gataggctga ctttgcacct cgttctcgta gcagaacacg ttgatgtcga tctggtccag
     1861 ggggatgtac tcgccctcgc tggtgatcct gctcttcttg tacagctcgt ccatgcccat
     1921 cacatcggta aaggcctttt gccactcctt gaagttgagc tcggtcttgg agtgcttgag
     1981 ctccgtctta cggaacacgt acatcggctg gttcttcaga tagttagccg ccattggctt
     2041 ggcaaaggtg taggtggtcc gcgcagtgct cctgtagcgc ttgccatttc cagtggtgta
     2101 actccactta aaggtactga tgatggtttt gtcgttgggg taagtcttct tcgacctgca
     2161 ccagtccgca gcggtcagcg agttggtcat cacaggaccg tcagcaggga aaccagtccc
     2221 cttcacctgg gcctctcctt tgatgtggct tccctcgtag gtgtagcggt agttaacagt
     2281 aagggaggca ccatcttcaa actgcattgt gcgatggact tggtatccgg agccatctac
     2341 catggcggcc tggaaaggcg acatcccgtc agggtagggc aggtactgat ggaagccata
     2401 cccgatatga gggaccagaa tccaggggga gaactggagg tcacccttgg tggacttcag
     2461 gtttaactcc tcataaccat catttggatt gccggtgccc tgacccacca tgtcaaagtc
     2521 cacaccgttg atggagccaa agatgtgtaa ctcatgtgtc gctgggtagg acttgaagat
     2581 agtctcatgg gcaatgaggc accccattcc aaacagtggc ttagcgagca catccaggat
     2641 tcccatgaga atggcctcat tgtcgacgga cagcacgcca ctaccagacc cgaaagccca
     2701 cacgacagga taaaacacga agaggattga caggaagacg agcagactga cataggagct
     2761 atggtactcg gggcttttct tcagatcgtc tttccaagtg gcgttataca tcccgtatgt
     2821 caggactgcc tgcatcacac atccagcgat aaagtacacc catttgaact tgtgttcatc
     2881 ctcgaaggcg cccagaatac caaacacgat catgaaaatg tctgcgccga tgacccaaga
     2941 aatcatcact ccgcccatct tggtcatgac gatgaggttg agcaggagca gtggtgtggt
     3001 cagcacccag tcaatatact ttgcgtagac cacctgcctt tcaccattca ttgtcaggtc
     3061 ctggtagttc acggccattg catagtaagc ggtagcggcg atagtgcaga tggcaatgtt
     3121 gatatagtat gcgctcttca gctctgaagg ccgaaagtgc acagccactc caaaagcaat
     3181 agcagcagca gccataatag caaagacaat ccactgtgca atcatgccgg tctcggtttc
     3241 cacgtcagcc atggtggcta gcgaagttcc tatactttct agagaatagg aacttcggaa
     3301 taggaacttc cgttgggatt cttcctattt tgatccaagc atcaccatcg accctctagt
     3361 ccagatctca ccatcgaccc gaagttccta taccttttga agaataggaa cttcggaata
     3421 ggaacttcaa tccccgcaat aagctcccat caagcttatc gataatcaac ctctggatta
     3481 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3541 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3601 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3661 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3721 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3781 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3841 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg
     3901 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3961 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     4021 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc
     4081 tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     4141 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     4201 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     4261 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     4321 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     4381 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     4441 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     4501 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     4561 ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     4621 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4681 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     4741 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     4801 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4861 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     4921 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4981 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     5041 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5101 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     5161 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     5221 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     5281 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     5341 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     5401 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     5461 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     5521 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5581 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5641 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5701 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5761 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5821 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5881 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5941 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6001 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6061 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6121 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6181 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6241 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6301 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6361 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6421 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6481 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6541 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6601 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6661 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6721 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6781 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6841 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6901 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6961 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7021 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7081 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7141 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7201 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7261 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7321 gccttttgct cacatgt
//
