LOCUS       sequence-417954-        6415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV viral vector packaging plasmid that expresses 
            tamoxifen-regulated Cre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6415)
  TITLE     Harvey/Richie Labs  AAV Cre plasmids
REFERENCE   2  (bases 1 to 6415)
  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: pOttc438 - pAAV EF1a CRE ERT2 bGHpA
FEATURES             Location/Qualifiers
     source          1..6415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        143..1321
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          374..1312
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1269..1289
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1344..2372
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVEDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             2385..3320
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2433..3326
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     primer_bind     complement(3353..3370)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3359..3470
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3590..3730
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3590..3719
                     /label=AAV2 ITR
     rep_origin      3805..4260
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3892..3911)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4102..4123
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4277..4296)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4396..4418
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4456..4474)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4542..4646
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4647..4715,4716..5507)
                     /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(4865..4884)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5678..6266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6167..6186
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6328..6415,1..42)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtaagc
       61 tttgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta
      121 ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc
      181 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
      241 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
      301 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
      361 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
      421 tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga
      481 gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc
      541 tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca
      601 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc
      661 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac
      721 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
      781 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
      841 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
      901 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
      961 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
     1021 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     1081 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg
     1141 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag
     1201 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     1261 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg
     1321 aggtaccgga tcctctcgac accatgtcca atttactgac cgtacaccaa aatttgcctg
     1381 cattaccggt cgatgcaacg agtgatgagg ttcgcaagaa cctgatggac atgttcaggg
     1441 atcgccaggc gttttctgag catacctgga aaatgcttct gtccgtttgc cggtcgtggg
     1501 cggcatggtg caagttgaat aaccggaaat ggtttcccgc agaacctgaa gatgttcgcg
     1561 attatcttct atatcttcag gcgcgcggtc tggcagtaaa aactatccag caacatttgg
     1621 gccagctaaa catgcttcat cgtcggtccg ggctgccacg accaagtgac agcaatgctg
     1681 tttcactggt tatgcggcgg atccgaaaag aaaacgttga tgccggtgaa cgtgcaaaac
     1741 aggctctagc gttcgaacgc actgatttcg accaggttcg ttcactcatg gaaaatagcg
     1801 atcgctgcca ggatatacgt aatctggcat ttctggggat tgcttataac accctgttac
     1861 gtatagccga aattgccagg atcagggttg aagatatctc acgtactgac ggtgggagaa
     1921 tgttaatcca tattggcaga acgaaaacgc tggttagcac cgcaggtgta gagaaggcac
     1981 ttagcctggg ggtaactaaa ctggtcgagc gatggatttc cgtctctggt gtagctgatg
     2041 atccgaataa ctacctgttt tgccgggtca gaaaaaatgg tgttgccgcg ccatctgcca
     2101 ccagccagct atcaactcgc gccctggaag ggatttttga agcaactcat cgattgattt
     2161 acggcgctaa ggatgactct ggtcagagat acctggcctg gtctggacac agtgcccgtg
     2221 tcggagccgc gcgagatatg gcccgcgctg gagtttcaat accggagatc atgcaagctg
     2281 gtggctggac caatgtaaat attgtcatga actatatccg taacctggat agtgaaacag
     2341 gggcaatggt gcgcctgctg gaagatggcg atctcgagcc atctgctgga gacatgagag
     2401 ctgccaacct ttggccaagc ccgctcatga tcaaacgctc taagaagaac agcctggcct
     2461 tgtccctgac ggccgaccag atggtcagtg ccttgttgga tgctgagccc cccatactct
     2521 attccgagta tgatcctacc agacccttca gtgaagcttc gatgatgggc ttactgacca
     2581 acctggcaga cagggagctg gttcacatga tcaactgggc gaagagggtg ccaggctttg
     2641 tggatttgac cctccatgat caggtccacc ttcttgaatg tgcctggcta gagatcctga
     2701 tgattggtct cgtctggcgc tccatggagc acccagtgaa gctactgttt gctcctaact
     2761 tgctcttgga caggaaccag ggaaaatgtg tagagggcat ggtggagatc ttcgacatgc
     2821 tgctggctac atcatctcgg ttccgcatga tgaatctgca gggagaggag tttgtgtgcc
     2881 tcaaatctat tattttgctt aattctggag tgtacacatt tctgtccagc accctgaagt
     2941 ctctggaaga gaaggaccat atccaccgag tcctggacaa gatcacagac actttgatcc
     3001 acctgatggc caaggcaggc ctgaccctgc agcagcagca ccagcggctg gcccagctcc
     3061 tcctcatcct ctcccacatc aggcacatga gtaacaaagg catggagcat ctgtacagca
     3121 tgaagtgcaa gaacgtggtg cccctctatg acctgctgct ggaggcggcg gacgcccacc
     3181 gcctacatgc gcccactagc cgtggagggg catccgtgga ggagacggac caaagccact
     3241 tggccactgc gggctctact tcatcgcatt ccttgcaaaa gtattacatc acgggggagg
     3301 cagagggttt ccctgccaca gcttaacgaa ttctaccggg taggagcgct agcctcgact
     3361 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3421 gaaggtgcca ctcccaccgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3481 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3541 gaagacaata gcaggcatgc tggggtacac gtgcggaccg agcggccgca ggaaccccta
     3601 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3661 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3721 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3781 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3841 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     3901 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3961 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4021 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4081 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     4141 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     4201 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     4261 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     4321 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     4381 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4441 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4501 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4561 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4621 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4681 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4741 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4801 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4861 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4921 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4981 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5041 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5101 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5161 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5221 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5281 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5341 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5401 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5461 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5521 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5581 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5641 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5701 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5761 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5821 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5881 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5941 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6001 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6061 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6121 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6181 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6241 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6301 ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg
     6361 aggccgcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgag
//
