LOCUS       Exported                6330 bp ds-DNA     circular SYN 24-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAAV_TRE_P68_HA_RINGnoNLS_TPR3_PIR_P2A_GFPii
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6330)
  AUTHORS   Don Arnold
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 13, 2025 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6330
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="AAV2 ITR"
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     promoter        159..473
                     /label=tight TRE promoter
                     /note="tight TRE promoter"
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    167..185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    203..221
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    238..256
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    274..292
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    310..328
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    345..363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    381..399
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    493..753
                     /note="PSD95.FiNGR"
     misc_feature    760..786
                     /note="HA tag"
     CDS             802..927
                     /codon_start=1
                     /note="Mdm2 RING"
                     /translation="CVICQGRPKNGCIVHGKTGHLMSCFTCAKALAAANAPCPVCR"
     CDS             886..909
                     /codon_start=1
                     /note="NLS mutated"
                     /translation="KALAAANA"
     misc_feature    934..1356
                     /note="TPR3"
     CDS             1363..1662
                     /codon_start=1
                     /note="PIR"
                     /translation="KKKLSKSDIMLVQSANVPSNPTQVPVEESGGFGSHHHNQNYCYQV
                     CLTPESAKTDLMFLKPCSPSRSTDAEHNPCGAIVTGYSDQQPDIISNGSILSNEN"
     misc_feature    1678..1734
                     /note="P2A"
     misc_feature    1744..2456
                     /note="eGFP"
     misc_feature    2460..3068
                     /label=WPRE
                     /note="WPRE"
     polyA_signal    3077..3553
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3593..3733
                     /label=AAV2 ITR
                     /note="AAV2 ITR"
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3808..4263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4545..4649
                     /gene="bla"
                     /label=AmpR promoter
                     /note="bla promoter"
     CDS             4650..5510
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5681..6269
                     /direction=RIGHT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtctcga gtttactccc tatcagtgat
      181 agagaacgta tgtcgagttt actccctatc agtgatagag aacgatgtcg agtttactcc
      241 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      301 cgagtttact ccctatcagt gatagagaac gtatgtcgag tttatcccta tcagtgatag
      361 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg
      421 tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagaa
      481 ttctctagaa tgctcgaagt caaggaagca tcaccaacca gcatccagat cagctgggtg
      541 ctccacttgc gccacgttcg ctactaccgc atcacctacg gtgaaactgg tggcaatagc
      601 cctgtccagg aattcaccgt gcctggcagc aagtccactg ctaccatcag cggcctgaaa
      661 cctggtgtcg actataccat cacggtgtac gccgtcacga tcttcagcgc ctaccgctcc
      721 gcctggccgc cgatctccat caactaccgc accaccggtt acccatacga tgttccagat
      781 tacgctggtg gaagcacgcg ttgtgtgatt tgccaggggc ggcctaaaaa tggttgcatt
      841 gttcacggca aaaccggaca cctgatgtca tgtttcacgt gtgcaaaggc actagctgcg
      901 gcgaatgcgc cctgccccgt gtgcagagct agcaagctct caggtggcgg agggagcggc
      961 ggaggtggct ctggaggtgg tggctcagct gaagcatggt acaatctggg aaacgcttat
     1021 tacaagcagg gcgactatca aaaagccatt gagtattacc aaaaggctct tgagctagac
     1081 cctaataacg ccgaagcgtg gtataaccta ggcaacgcat attacaagca gggagactac
     1141 caaaaggcaa tcgaatatta ccagaaggca ctcgaactgg acccaaataa cgcagaggct
     1201 tggtataatc tgggaaacgc ctactataaa cagggtgact accaaaaggc gatagaggac
     1261 tatcagaagg ccttggagct cgatccgaac aatctgcaac gtagtgctgg tggcggaggg
     1321 tctggtggag gaggctcagg aggaggtggc gcatcaggta ccaagaagaa actcagcaag
     1381 tcagacatca tgttggtgca gagcgccaat gtccctagca acccgaccca ggtgccggta
     1441 gaggagtccg gaggcttcgg ttcccatcac cacaatcaga actattgcta tcaggtctgc
     1501 ctcaccccag agtccgccaa gaccgacctg atgttcttga agccctgcag cccttcccgg
     1561 agtacagacg ctgagcacaa tccctgtgga gccatcgtca ccgggtacag cgaccagcaa
     1621 ccagacatca tttccaacgg aagcattttg tccaacgaga atggtggatc atgtacagct
     1681 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctggaggt
     1741 actggggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1801 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1861 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1921 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     1981 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2041 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2101 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2161 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     2221 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2281 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2341 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     2401 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtaaaag
     2461 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2521 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2581 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2641 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2701 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2761 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2821 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     2881 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2941 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3001 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3061 cgataccgag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     3121 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     3181 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     3241 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     3301 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     3361 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     3421 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     3481 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     3541 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     3601 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     3661 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     3721 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3781 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     3841 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3901 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3961 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4021 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4081 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4141 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4201 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     4261 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4321 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4381 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4441 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4501 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4561 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4621 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4681 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4741 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4801 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4861 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4921 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4981 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5041 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5101 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5161 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5221 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5281 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5341 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5401 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5461 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5521 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5581 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5641 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5701 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5761 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5821 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5881 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5941 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6001 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6061 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6121 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6181 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6241 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6301 tggccttttg ctggcctttt gctcacatgt
//
