LOCUS       Exported                5441 bp DNA     circular SYN 19-MAY-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-TreTight2-Akaluc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5441)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 19, 2022 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
COMMENT     ORIGDB|GenBank
FEATURES             Location/Qualifiers
     source          1..5441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     protein_bind    175..196
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        211..241
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 211 .. 216 / #ffffff / -35
                        2: 217 .. 234 / #ffffff
                        3: 235 .. 241 / #ffffff / -10"
     misc_feature    249..498
                     /label=TREmod
                     /note="/vntifkey=21"
                     /note="color: #66cc99; direction: RIGHT"
     promoter        250..564
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    258..276
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    294..312
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    329..347
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    365..383
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    401..419
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    436..454
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    472..490
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     regulatory      596..605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     misc_feature    602..2254
                     /label=Akaluc
                     /note="/vntifkey=21"
                     /note="color: #66cc99; direction: RIGHT"
     repeat_region   2704..2844
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      2919..3374
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3656..3760
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3761..4621
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3761 .. 3829 / #ccffcc / signal sequence
                        2: 3830 .. 4621 / #ccffcc
                     Cleavage site after base 3829"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4792..5380
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca gatgctgcac acttgtgggc tagctaatgt
      181 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      241 gtgtggctcg agtttactcc ctatcagtga tagagaacgt atgtcgagtt tactccctat
      301 cagtgataga gaacgatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagt
      361 ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa
      421 cgtatgtcga gtttatccct atcagtgata gagaacgtat gtcgagttta ctccctatca
      481 gtgatagaga acgtatgtcg aggtaggcgt gtacggtggg aggcctatat aagcagagct
      541 cgtttagtga accgtcagat cgcctggaga attcgagctc ggtacccggg gatccgccac
      601 catggaagat gccaaaaaca ttaagaaggg cccagcgccg ttctacccac tcgaagacgg
      661 gaccgccggc gagcagctgc acaaagccat gaagcgctac gccctggtgc ccggcgccat
      721 cgcctttacc gacgcacata ttcaggtgga cgttacctac gccgagtact tcgagatgag
      781 cgttcggctg gcagaagcta tgaggcgcta tgggctgaat acaaaccatc ggatcgtggt
      841 gtgcagcgag aatagctcgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt
      901 ggctgtggcc ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat
      961 cagccagccc accgtcgtat tcgtgagcaa gaaagggctg cgaaaggtcc tcaacgtgca
     1021 aaagaagcta ccgatcatac gaaagatcat catcatggat agcaagaccg actaccaggg
     1081 cttccaaagc atgtacacct tcgtgacttc ccatttgcca cccagcttca acgagtacga
     1141 cttcgtgccc gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg
     1201 tagtacagga ttaccaaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag
     1261 tcatgcccgc gaccccatct tcggctacca gaacatcccc gacaccgcta tcctcagcgt
     1321 ggtgccattt caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt
     1381 tcgggtcgtg ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta
     1441 taagattcaa tctgccctgc tggtgcccac actatttagc tgcctcgcta agagcactct
     1501 catcgacaag tacgacctaa gcagcttgcg cgagatcgcc agcggcgggg cgccgctcag
     1561 caaggaggta ggtgaggccg tggccaaacg cttccgccta ccaggcatcc gccagggcta
     1621 tggcctgaca gaaacaacca acgccgtcat gatcaccccc gagggggacc gtaagcctgg
     1681 ctcagtaggc aaggtggtgc ccttcttcga ggctaaggtg gtagacttgg tcaccggtaa
     1741 gacactgggt gtgaaccagc gcggtgagct gtgcgtccgt ggccccatga tcatgagcgg
     1801 ctacgttaac aaccccgagg ctacgaacgc tctcatcgac aaggacggct ggctgcacag
     1861 cggcgacatc gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag
     1921 cctgatcaaa tacaagggct accaggtagc cccagccgaa ctggagggca tcctgctgca
     1981 acacccctac atcttcgacg ccggagtcgc cggcctgccc gacgacgatg ccggcgagct
     2041 gcccgccgca gtcgtcgtgt tggaacacgg taaaaccatg accgagaaag agatcgtgga
     2101 ctatgtggcc agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgtttgtgga
     2161 tgaagtccct agaggatcga ccggcaagtt agacgcccgc aagatccgcg agattctcac
     2221 taaggccaag aaggacggca agatcgccgt gtaagaattc cgctcgagat aatcaacctc
     2281 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2341 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2401 ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct
     2461 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttta
     2521 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat ctagctttat ttgtgaaatt
     2581 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2641 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcggc
     2701 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2761 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2821 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     2881 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     2941 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3001 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3061 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3121 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3181 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3241 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     3301 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3361 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3421 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3481 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3541 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3601 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3661 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3721 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3781 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3841 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3901 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3961 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4021 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4081 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4141 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4201 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4261 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4321 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4381 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4441 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4501 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4561 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4621 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4681 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4741 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4801 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4861 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4921 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4981 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5041 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5101 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5161 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5221 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5281 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5341 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5401 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
