LOCUS       sequence-463794-        6925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To Express a CRE dependent Flag tagged Mouse Beta Adrenergic 
            Receptor from a EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6925)
  TITLE     Ploski Lab Plasmids 2025
REFERENCE   2  (bases 1 to 6925)
  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"
COMMENT     Sequence Label: pAAV-EF1a-DIO-FLAG-mouse Beta Adrenergic Recep...
FEATURES             Location/Qualifiers
     source          1..6925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        881..2059
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1112..2050
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2007..2027
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(2090..2123)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2174..2207)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2507..2518
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3560..3593
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3644..3677
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3702..4290
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3755..3775)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4173..4184)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4322..4798
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4521..4540)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4838..4978
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4838..4967
                     /label=AAV2 ITR
     rep_origin      5053..5508
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5140..5159)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5350..5371
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5525..5544)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5644..5666
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5704..5722)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5790..5894
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5895..5963,5964..6755)
                     /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(6113..6132)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg
      841 cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg
      901 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      961 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     1021 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     1081 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
     1141 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta
     1201 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
     1261 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
     1321 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
     1381 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
     1441 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
     1501 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     1561 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     1621 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     1681 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     1741 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     1801 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     1861 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     1921 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     1981 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     2041 tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa taacttcgta
     2101 taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc tattagcaat
     2161 atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc gcgccttaca
     2221 gtggcgagtc atttgtacta cagttccttc cttgggagtc aacgctaagg ctaggcacag
     2281 taccttgaca gttcacaaag ccttccatgc ctgggggatc ctcacacagc agttcctgtt
     2341 ctctctcctg ccccagctga caagtgtttg gctcccctgt gtagtccgtt ctgccgttgc
     2401 tattgctaga gtagccgttc ccataggttt tcgaagaaga cctgcgaagg cacagaagct
     2461 cttgaaaggc aatcctgaaa tctggactcc gacagtagat aagaggattg aaggcagagt
     2521 tgacgtagcc caaccagtta aggaggatgt aaacttcctt agggatgagg ttgtccctga
     2581 taacgtgcac gatattgaca atgaagaagg gcagccagca gagggtgaat gtgcccatga
     2641 tgatgcctaa agtcttgagg gctttgtgct ctttcaagca gaacttggag gaccttcgga
     2701 gtccgtggcc gctccgccca tcctgctcca cctggctgag gttttgggcg tggaatcttc
     2761 cttcagattt gtctatcttc tgcagctgcc ttttggccac ctggaagacc cgggaataga
     2821 caaagaccat caccaccagg ggcacgtaga aagacacaat cgaggacgcg atggcgtagg
     2881 cctggttcgt gaagaagtca cagcaagtct cctcggtgta acaatcgata gctttcttgt
     2941 gggtggcacg gtaccagtgc atctggatag gcaaaaagga ggtaaggcca gatacaatcc
     3001 ataccatcag gatgaccact cgggccttat tcttggtcag caggctctgg tacttgaagg
     3061 gcgatgtgat agcaacatag cgatccactg caatcacgca cagggtctcg atgctggctg
     3121 tgacgcacaa cacatcaatg gaagtccaga actcgcacca gaagttgcca aaattccaca
     3181 ttttcataag gatgtgactg gccccaaacg gcaccaccgc taggcccatg actagatcag
     3241 cacacgccaa ggagattatg aagtagttgg tgacggtttg tagtcgctcg aacttggcaa
     3301 tggctgtgat gaccagcacg ttgccaaaca cgatggccag gacgataacc gacatgagga
     3361 tggccatgcc cacaacccac gcttcgtccc gttcctgagt gacgtcgtgg tctggcgctc
     3421 ggcttccgtt gggtgccagc aagaagtcgc tgtcgttccc gtgtggcccc atggcgtcat
     3481 catcgtcctt gtagtcggcg aacaccaggc agaagatgta gctcagggcg atgatggtct
     3541 tcatggtggc ggcgctagca taacttcgta taaagtatcc tatacgaagt tatttgcctt
     3601 aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt cgtataatgt
     3661 atgctatacg aagttatgaa ttcgatatca agcttatcga taatcaacct ctggattaca
     3721 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3781 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3841 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3901 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3961 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     4021 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     4081 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     4141 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     4201 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     4261 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta
     4321 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     4381 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     4441 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     4501 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     4561 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     4621 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     4681 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     4741 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     4801 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     4861 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     4921 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     4981 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     5041 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5101 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     5161 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5221 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     5281 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5341 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     5401 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     5461 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     5521 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     5581 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5641 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     5701 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5761 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5821 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5881 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5941 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6001 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6061 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6121 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6181 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6241 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6301 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6361 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6421 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6481 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6541 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6601 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6661 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6721 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6781 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6841 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6901 ccgtagaaaa gatcaaagga tcttc
//
