LOCUS       Exported                8569 bp ds-DNA     circular SYN 05-APR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8569)
  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..8569
                     /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"
     primer_bind     1542..1579
                     /label=VLDLR_fwd
     CDS             1562..3886
                     /codon_start=1
                     /label=VLDLR(1-979)_751-779->R
                     /translation="MGTSALWALWLLLALCWAPRESGATGTGRKAKCEPSQFQCTNGRC
                     ITLLWKCDGDEDCVDGSDEKNCVKKTCAESDFVCNNGQCVPSRWKCDGDPDCEDGSDES
                     PEQCHMRTCRIHEISCGAHSTQCIPVSWRCDGENDCDSGEDEENCGNITCSPDEFTCSS
                     GRCISRNFVCNGQDDCSDGSDELDCAPPTCGAHEFQCSTSSCIPISWVCDDDADCSDQS
                     DESLEQCGRQPVIHTKCPASEIQCGSGECIHKKWRCDGDPDCKDGSDEVNCPSRTCRPD
                     QFECEDGSCIHGSRQCNGIRDCVDGSDEVNCKNVNQCLGPGKFKCRSGECIDISKVCNQ
                     EQDCRDWSDEPLKECHINECLVNNGGCSHICKDLVIGYECDCAAGFELIDRKTCGDIDE
                     CQNPGICSQICINLKGGYKCECSRGYQMDLATGVCKAVGKEPSLIFTNRRDIRKIGLER
                     KEYIQLVEQLRNTVALDADIAAQKLFWADLSQKAIFSASIDDKVGRHVKMIDNVYNPAA
                     IAVDWVYKTIYWTDAASKTISVATLDGTKRKFLFNSDLREPASIAVDPLSGFVYWSDWG
                     EPAKIEKAGMNGFDRRPLVTADIQWPNGITLDLIKSRLYWLDSKLHMLSSVDLNGQDRR
                     IVLKSLEFLAHPLALTIFEDRVYWIDGENEAVYGANKFTGSELATLVNNLNDAQDIIVY
                     HELVQPSGKNWCEEDMENGGCEYLCLPAPQINDHSPKYTCSCPSGYNVEENGRDCQRIN
                     VTTAVSEVSVPPKGTSGEPEA"
     primer_bind     1643..1661
                     /label=TIM371
     primer_bind     2312..2341
                     /label=VLDLR_750_seq
     primer_bind     complement(2607..2627)
                     /label=TIM372
     primer_bind     complement(3830..3868)
                     /label=VLDLR_rev_inner
     primer_bind     complement(3841..3888)
                     /label=VLDLR_rev_inner2
     primer_bind     complement(3861..3906)
                     /label=VLDLR-EPEA_rev_outer
     primer_bind     complement(3974..3991)
                     /label=BGH-rev
     polyA_signal    3980..4204
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    4488..4535
                     /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      4629..4763
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4768..5370
                     /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    5604..5725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(6378..6396)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6417..6433)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6441..6457
                     /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(6465..6495)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6510..6531
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6819..7407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7578..8438)
                     /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(8439..8543)
                     /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 gatgggcacg tccgcgctct gggcgctctg gctgctgctc gcgctgtgct gggcgccccg
     1621 ggagagcggc gccaccggaa ccgggagaaa agccaaatgt gaaccctccc aattccagtg
     1681 cacaaatggt cgctgtatta cgctgttgtg gaaatgtgat ggggatgaag actgtgttga
     1741 cggcagtgat gaaaagaact gtgtaaagaa gacgtgtgct gaatctgact tcgtgtgcaa
     1801 caatggccag tgtgttccca gccgatggaa gtgtgatgga gatcctgact gcgaagatgg
     1861 ttcagatgaa agcccagaac agtgccatat gagaacatgc cgcatacatg aaatcagctg
     1921 tggcgcccat tctactcagt gtatcccagt gtcctggaga tgtgatggtg aaaatgattg
     1981 tgacagtgga gaagatgaag aaaactgtgg caatataaca tgtagtcccg acgagttcac
     2041 ctgctccagt ggccgctgca tctccaggaa ctttgtatgc aatggccagg atgactgcag
     2101 cgatggcagt gatgagctgg actgtgcccc gccaacctgt ggcgcccatg agttccagtg
     2161 cagcacctcc tcctgcatcc ccatcagctg ggtatgcgac gatgatgcag actgctccga
     2221 ccaatctgat gagtccctgg agcagtgtgg ccgtcagcca gtcatacaca ccaagtgtcc
     2281 agccagcgaa atccagtgcg gctctggcga gtgcatccat aagaagtggc gatgtgatgg
     2341 ggaccctgac tgcaaggatg gcagtgatga ggtcaactgt ccctctcgaa cttgccgacc
     2401 tgaccaattt gaatgtgagg atggcagctg catccatggc agcaggcagt gtaatggtat
     2461 ccgagactgt gtcgatggtt ccgatgaagt caactgcaaa aatgtcaatc agtgcttggg
     2521 ccctggaaaa ttcaagtgca gaagtggaga atgcatagat atcagcaaag tatgtaacca
     2581 ggagcaggac tgcagggact ggagtgatga gcccctgaaa gagtgtcata taaacgaatg
     2641 cttggtaaat aatggtggat gttctcatat ctgcaaagac ctagttatag gctacgagtg
     2701 tgactgtgca gctgggtttg aactgataga taggaaaacc tgtggagata ttgatgaatg
     2761 ccaaaatcca ggaatctgca gtcaaatttg tatcaactta aaaggcggtt acaagtgtga
     2821 atgtagtcgt ggctatcaaa tggatcttgc tactggcgtg tgcaaggcag taggcaaaga
     2881 gccaagtctg atcttcacta atcgaagaga catcaggaag attggcttag agaggaaaga
     2941 atatatccaa ctagttgaac agctaagaaa cactgtggct ctcgatgctg acattgctgc
     3001 ccagaaacta ttctgggccg atctaagcca aaaggctatc ttcagtgcct caattgatga
     3061 caaggttggt agacatgtta aaatgatcga caatgtctat aatcctgcag ccattgctgt
     3121 tgattgggtg tacaagacca tctactggac tgatgcggct tctaagacta tttcagtagc
     3181 taccctagat ggaaccaaga ggaagttcct gtttaactct gacttgcgag agcctgcctc
     3241 catagctgtg gacccactgt ctggctttgt ttactggtca gactggggtg aaccagctaa
     3301 aatagaaaaa gcaggaatga atggattcga tagacgtcca ctggtgacag cggatatcca
     3361 gtggcctaac ggaattacac ttgaccttat aaaaagtcgc ctctattggc ttgattctaa
     3421 gttgcacatg ttatccagcg tggacttgaa tggccaagat cgtaggatag tactaaagtc
     3481 tctggagttc ctagctcatc ctcttgcact aacaatattt gaggatcgtg tctactggat
     3541 agatggggaa aatgaagcag tctatggtgc caataaattc actggatcag agctagccac
     3601 tctagtcaac aacctgaatg atgcccagga catcattgtc tatcatgaac ttgtacagcc
     3661 atcaggtaaa aattggtgtg aagaagacat ggagaatgga ggatgtgaat acctatgcct
     3721 gccagcacca cagattaatg atcactctcc aaaatatacc tgttcctgtc ccagtgggta
     3781 caatgtagag gaaaatggcc gagactgtca aaggatcaat gtgaccacag cagtatcaga
     3841 ggtcagtgtt cccccaaaag ggacttctgg agaaccagaa gcctaagcat agataactga
     3901 tccagtgtgc tgggctacga tgcatgggcc ccctaggtcg agtctagagg gcccgtttaa
     3961 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4021 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4081 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4141 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4201 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     4261 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4321 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4381 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4441 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa gttcctattc
     4501 cgaagttcct attctctaga aagtatagga acttccttgg ccaaaaagac cggtaccagg
     4561 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4621 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4681 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4741 agtgaggagg cttttttgga ggctaccatg ggtaccgagt acaagcccac ggtgcgcctc
     4801 gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4861 cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa
     4921 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     4981 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5041 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5101 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     5161 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5221 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5281 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagt gcccgaagga ccgcgcgacc
     5341 tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc
     5401 gaaaggagcg cacgacccca tggctccgac cgaagccgac ccgggcggcc ccgccgaccc
     5461 cgcacccgcc cccgaggccc accgactcta gataactgat cataatcagc cataccacat
     5521 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5581 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5641 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5701 tgtccaaact catcaatgta tcttaacgcg tcgagtgcat tctagttgtg gtttgtccaa
     5761 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctgca ggaattcgat
     5821 atcaagctct agccaacaag ctcgtcatcg ctttgcagaa gagcagagag gatatgctca
     5881 tcgtctaaag aactacccat tttattatat attagtcacg atatctataa caagaaaata
     5941 tatatataat aagttatcac gtaagtagaa catgaaataa caatataatt atcgtatgag
     6001 ttaaatctta aaagtcacgt aaaagataat catgcgtcat tttgactcac gcggtcgtta
     6061 tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg cctcacggga gctccaagcg
     6121 gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa gagagagcaa
     6181 tatttcaaga atgcatgcgt caattttacg cagactatct ttctagggtt aaaaaagatt
     6241 tgcgctttac tcgacctaaa ctttaaacac gtcatagaat cttcgtttga caaaaaccac
     6301 attgtggcca agctgtgtga cgcgacgcgc gctaaagaat ggcaaaccaa gtcgcgcgag
     6361 gtacccagct tttgttccct ttagtgaggg ttaattccga gcttggcgta atcatggtca
     6421 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6481 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6541 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6601 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6661 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6721 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6781 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6841 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6901 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6961 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7021 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7081 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7141 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7201 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     7261 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7321 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7381 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7441 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7501 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7561 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7621 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7681 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7741 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7801 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7861 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7921 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7981 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8041 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8101 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8161 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8221 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8281 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8341 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8401 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8461 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8521 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
