LOCUS       Exported                7534 bp ds-DNA     circular SYN 19-JAN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7534)
  AUTHORS   MPIB
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 19, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7534
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1145..1163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1182..1200
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1219..1237
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1256..1274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1293..1311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1330..1348
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1367..1385
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1404..1422
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1571..2842
                     /codon_start=1
                     /label=Clusterin
                     /translation="MMKTLLLFVGLLLTWESGQVLGDQTVSDNELQEMSNQGSKYVNKE
                     IQNAVNGVKQIKTLIEKTNEERKTLLSNLEEAKKKKEDALNETRESETKLKELPGVCNE
                     TMMALWEECKPCLKQTCMKFYARVCRSGSGLVGRQLEEFLNQSSPFYFWMNGDRIDSLL
                     ENDRQQTHMLDVMQDHFSRASSIIDELFQDRFFTREPQDTYHYLPFSLPHNFHAMFQPF
                     LEMIHEAQQAMDIHFHSPAFQHPPTEFIREGDDDRTVCREIRHNSTGCLRMKDQCDKCR
                     EILSVDCSTNNPSQAKLRRELDESLQVAERLTRKYNELLKSYQWKMLNTSSLLEQLNEQ
                     FNWVSRLANLTQGEDQYYLRVTTVASHTSDSDVPSGVTEVVVKLFDSDPITVTVPVEVS
                     RKNPKFMETVAEKALQEYRKKHREE"
     misc_feature    1571..1636
                     /label=signal peptide
     polyA_signal    2945..3169
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3453..3500
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     rep_origin      3594..3728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3733..4335
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     polyA_signal    4569..4690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(5343..5361)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5382..5398)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5406..5422
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5430..5460)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5475..5496
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5784..6372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6543..7403)
                     /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"
     promoter        complement(7404..7508)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcgg
      661 tattcacgac agcaggctga ataataaaaa aattagaaac tattatttaa ccctagaaag
      721 ataatcatat tgtgacgtac gttaaagata atcatgcgta aaattgacgc atgtgtttta
      781 tcggtctgta tatcgaggtt tatttattaa tttgaataga tattaagttt tattatattt
      841 acacttacat actaataata aattcaacaa acaatttatt tatgtttatt tatttattaa
      901 aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa aacttttaaa cattctctct
      961 tttacaaaaa taaacttatt ttgtacttta aaaacagtca tgttgtatta taaaataagt
     1021 aattagctta acctatacat aatagaaaca aattatactt attagtcagt cagaaacaac
     1081 tttggcacat atcaatatta tgctctcgtt aattaactcg agggatccaa gctggccgca
     1141 cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca gtgatagaga
     1201 aggtacgtct agaacgtctc cctatcagtg atagagaagt cgacacgtct cgagctccct
     1261 atcagtgata gagaaggtac gtctagaacg tctccctatc agtgatagag aagtcgacac
     1321 gtctcgagct ccctatcagt gatagagaag gtacgtctag aacgtctccc tatcagtgat
     1381 agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtaccc cctatataag
     1441 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
     1501 ccatagaaga caccgggacc gatccagcct ccgcggtggc ggccatcaca agtttgtaca
     1561 gctagccacc atgatgaaga ctctgctgct gtttgtgggg ctgctgctga cctgggagag
     1621 tgggcaggtc ctgggggacc agacggtctc agacaatgag ctccaggaaa tgtccaatca
     1681 gggaagtaag tacgtcaata aggaaattca aaatgctgtc aacggggtga aacagataaa
     1741 gactctcata gaaaaaacaa acgaagagcg caagacactg ctcagcaacc tagaagaagc
     1801 caagaagaag aaagaggatg ccctaaatga gaccagggaa tcagagacaa agctgaagga
     1861 gctcccagga gtgtgcaatg agaccatgat ggccctctgg gaagagtgta agccctgcct
     1921 gaaacagacc tgcatgaagt tctacgcacg cgtctgcaga agtggctcag gcctggttgg
     1981 ccgccagctt gaggagttcc tgaaccagag ctcgcccttc tacttctgga tgaatggtga
     2041 ccgcatcgac tccctgctgg agaacgaccg gcagcagacg cacatgctgg atgtcatgca
     2101 ggaccacttc agccgcgcgt ccagcatcat agacgagctc ttccaggaca ggttcttcac
     2161 ccgggagccc caggatacct accactacct gcccttcagc ctgccccaca acttccacgc
     2221 catgttccag cccttccttg agatgataca cgaggctcag caggccatgg acatccactt
     2281 ccatagcccg gccttccagc acccgccaac agaattcata cgagaaggcg acgatgaccg
     2341 gactgtgtgc cgggagatcc gccacaactc cacgggctgc ctgcggatga aggaccagtg
     2401 tgacaagtgc cgggagatct tgtctgtgga ctgttccacc aacaacccct cccaggctaa
     2461 gctgcggcgg gagctcgacg aatccctcca ggtcgctgag aggttgacca ggaaatacaa
     2521 cgagctgcta aagtcctacc agtggaagat gctcaacacc tcctccttgc tggagcagct
     2581 gaacgagcag tttaactggg tgtcccggct ggcaaacctc acgcaaggcg aagaccagta
     2641 ctatctgcgg gtcaccacgg tggcttccca cacttctgac tcggacgttc cttccggtgt
     2701 cactgaggtg gtcgtgaagc tctttgactc tgatcccatc actgtgacgg tccctgtaga
     2761 agtctccagg aagaacccta aatttatgga gaccgtggcg gagaaagcgc tgcaggaata
     2821 ccgcaaaaag caccgggagg agtgagcggc cgcatagata actgatccag tgtgctgggc
     2881 tacgatgcat gggcccccta ggtcgagtct agagggcccg tttaaacccg ctgatcagcc
     2941 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3001 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3061 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3121 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3181 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3241 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3301 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3361 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3421 aaacttgatt agggtgatgg ttcacgtacc tagaagttcc tattccgaag ttcctattct
     3481 ctagaaagta taggaacttc cttggccaaa aagaccggta ccaggcaggc agaagtatgc
     3541 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3601 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3661 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3721 ttggaggcta ccatgggtac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     3781 gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     3841 accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     3901 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     3961 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     4021 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     4081 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     4141 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     4201 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     4261 ttcaccgtca ccgccgacgt cgagtgcccg aaggaccgcg cgacctggtg catgacccgc
     4321 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
     4381 ccccatggct ccgaccgaag ccgacccggg cggccccgcc gaccccgcac ccgcccccga
     4441 ggcccaccga ctctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     4501 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     4561 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4621 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4681 atgtatctta acgcgtcgag tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4741 ttatcatgtc tgtataccgt cgacctctag ctgcaggaat tcgatatcaa gctctagcca
     4801 acaagctcgt catcgctttg cagaagagca gagaggatat gctcatcgtc taaagaacta
     4861 cccattttat tatatattag tcacgatatc tataacaaga aaatatatat ataataagtt
     4921 atcacgtaag tagaacatga aataacaata taattatcgt atgagttaaa tcttaaaagt
     4981 cacgtaaaag ataatcatgc gtcattttga ctcacgcggt cgttatagtt caaaatcagt
     5041 gacacttacc gcattgacaa gcacgcctca cgggagctcc aagcggcgac tgagatgtcc
     5101 taaatgcaca gcgacggatt cgcgctattt agaaagagag agcaatattt caagaatgca
     5161 tgcgtcaatt ttacgcagac tatctttcta gggttaaaaa agatttgcgc tttactcgac
     5221 ctaaacttta aacacgtcat agaatcttcg tttgacaaaa accacattgt ggccaagctg
     5281 tgtgacgcga cgcgcgctaa agaatggcaa accaagtcgc gcgaggtacc cagcttttgt
     5341 tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5401 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5461 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5521 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5581 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5641 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5701 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5761 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5821 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5881 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5941 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6001 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6061 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6121 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6181 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6241 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6301 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6361 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6421 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6481 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6541 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6601 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6661 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6721 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6781 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6841 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6901 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6961 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7021 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7081 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7141 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7201 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7261 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7321 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7381 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7441 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7501 gttccgcgca catttccccg aaaagtgcca cctg
//
