LOCUS       Exported                5700 bp ds-DNA     circular SYN 31-JAN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    TRE-PFE3-HA final
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5700)
  AUTHORS   Don Arnold
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 7, 2025 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5700
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        30..408
                     /note="TRE3G promoter"
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    38..56
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    74..92
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    110..128
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    146..164
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    182..200
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    218..236
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    254..272
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    428..688
                     /note="PSD95.FiNGR"
     misc_feature    695..721
                     /note="HA tag"
     CDS             737..862
                     /codon_start=1
                     /note="Mdm2 RING"
                     /translation="CVICQGRPKNGCIVHGKTGHLMSCFTCAKALAAANAPCPVCR"
     CDS             821..844
                     /codon_start=1
                     /note="NLS mutated"
                     /translation="KALAAANA"
     misc_feature    869..1291
                     /note="TPR3"
     CDS             1298..1597
                     /codon_start=1
                     /note="PIR"
                     /translation="KKKLSKSDIMLVQSANVPSNPTQVPVEESGGFGSHHHNQNYCYQV
                     CLTPESAKTDLMFLKPCSPSRSTDAEHNPCGAIVTGYSDQQPDIISNGSILSNEN"
     intron          1623..1756
                     /note="chimeric intron"
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    1765..2353
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    2386..2497
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3950..4538)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4709..5569)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5570..5674)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 gtcgacattg attattgact agttatctcg agtttactcc ctatcagtga tagagaacgt
       61 atgaagagtt tactccctat cagtgataga gaacgtatgc agactttact ccctatcagt
      121 gatagagaac gtataaggag tttactccct atcagtgata gagaacgtat gaccagttta
      181 ctccctatca gtgatagaga acgtatctac agtttactcc ctatcagtga tagagaacgt
      241 atatccagtt tactccctat cagtgataga gaacgtataa gctttaggcg tgtacggtgg
      301 gcgcctataa aagcagagct cgtttagtga accgtcagat cgcctggagc aattccacaa
      361 cacttttgtc ttataccaac tttccgtacc acttcctacc ctcgtaaagt cgacaccggg
      421 gcccatgctc gaagtcaagg aagcatcacc aaccagcatc cagatcagct gggtgctcca
      481 cttgcgccac gttcgctact accgcatcac ctacggtgaa actggtggca atagccctgt
      541 ccaggaattc accgtgcctg gcagcaagtc cactgctacc atcagcggcc tgaaacctgg
      601 tgtcgactat accatcacgg tgtacgccgt cacgatcttc agcgcctacc gctccgcctg
      661 gccgccgatc tccatcaact accgcaccac cggttaccca tacgatgttc cagattacgc
      721 tggtggaagc acgcgttgtg tgatttgcca ggggcggcct aaaaatggtt gcattgttca
      781 cggcaaaacc ggacacctga tgtcatgttt cacgtgtgca aaggcactag ctgcggcgaa
      841 tgcgccctgc cccgtgtgca gagctagcaa gctctcaggt ggcggaggga gcggcggagg
      901 tggctctgga ggtggtggct cagctgaagc atggtacaat ctgggaaacg cttattacaa
      961 gcagggcgac tatcaaaaag ccattgagta ttaccaaaag gctcttgagc tagaccctaa
     1021 taacgccgaa gcgtggtata acctaggcaa cgcatattac aagcagggag actaccaaaa
     1081 ggcaatcgaa tattaccaga aggcactcga actggaccca aataacgcag aggcttggta
     1141 taatctggga aacgcctact ataaacaggg tgactaccaa aaggcgatag aggactatca
     1201 gaaggccttg gagctcgatc cgaacaatct gcaacgtagt gctggtggcg gagggtctgg
     1261 tggaggaggc tcaggaggag gtggcgcatc aggtaccaag aagaaactca gcaagtcaga
     1321 catcatgttg gtgcagagcg ccaatgtccc tagcaacccg acccaggtgc cggtagagga
     1381 gtccggaggc ttcggttccc atcaccacaa tcagaactat tgctatcagg tctgcctcac
     1441 cccagagtcc gccaagaccg acctgatgtt cttgaagccc tgcagccctt cccggagtac
     1501 agacgctgag cacaatccct gtggagccat cgtcaccggg tacagcgacc agcaaccaga
     1561 catcatttcc aacggaagca ttttgtccaa cgagaattaa atcgattatg tcgaggccgc
     1621 aggtaagtat caaggttaca agacaggttt aaggagacca catagaaact gggcttgtcg
     1681 agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt
     1741 gcctttctct ccacaggtgt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     1801 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     1861 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     1921 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     1981 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2041 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2101 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     2161 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2221 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2281 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2341 gccgcctccc cgcctggaat tcgagctcgg tacgatcagc ctcgactgtg ccttctagtt
     2401 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2461 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2521 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2581 ggccagcttt tgttcccttt agggttaatt gcgcgcttgg cgctaatcat ggtcatagct
     2641 gtttccactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     2701 ctggctcaca aataccactg agatcgtttt ccctctgcca aaaattatgg ggacatcatg
     2761 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     2821 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     2881 catcagaatg agtatttggt ttagagtttg gcaacatatg ccatatgctg gctgccatga
     2941 acaaaggtgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     3001 attccataga aaagccttga cttgaggtta gatttttttt tatattttgt ttgtgttatt
     3061 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     3121 ctcctgacta ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacctgca
     3181 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3241 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3301 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3361 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3421 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3481 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3541 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     3601 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3661 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
     3721 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3781 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3841 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3901 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3961 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4021 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4081 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4141 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4201 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4261 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4321 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4381 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4441 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4501 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4561 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4621 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4681 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4741 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4801 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4861 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4921 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4981 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5041 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5101 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5161 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5221 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5281 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5341 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5401 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5461 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5521 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5581 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5641 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
