LOCUS       pNtVimentin-mche        7221 bp    DNA     circular SYN 01-NOV-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7221)
  AUTHORS   Alessandra Affolter Lab
  TITLE     Direct Submission
  JOURNAL   Exported Jun 11, 2025 from SnapGene 8.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7221
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     primer_bind     223..263
                     /label=testcmvpromfor
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     720..740
                     /label=CMVfor
     primer_bind     720..740
                     /label=CMV-for
     promoter        731..769
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    771..789
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    792..810
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     945..973
                     /label=ntvimentinfor
     misc_feature    956..2350
                     /label=mmVimentin
                     /note="mmVimentin"
     primer_bind     complement(2330..2357)
                     /label=ntvimentinrev
     primer_bind     2351..2377
                     /label=mTFPmito_for
     CDS             2360..3064
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     2360..2377
                     /label=testtfprab5cmvfor
     primer_bind     2360..2374
                     /label=mcherry5pletm1_for
     primer_bind     2998..3013
                     /label=mcherry for
     primer_bind     complement(3036..3064)
                     /label=mtfprab5f.REV
     primer_bind     complement(3043..3063)
                     /label=testtfprabcmvrev
     primer_bind     complement(3046..3088)
                     /label=mTFPmito_rev
     primer_bind     complement(3050..3084)
                     /label=mcherry3pletm1_rev
     primer_bind     complement(3050..3084)
                     /label=mcherry3pletm1_reva
     primer_bind     3125..3153
                     /label=HAformito_for
     primer_bind     3136..3163
                     /label=testhabspe1for
     primer_bind     complement(3137..3164)
                     /label=testhahind3rev
     CDS             3137..3163
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3149..3180)
                     /label=HAformito_rev
     primer_bind     complement(3232..3249)
                     /label=BGH-rev
     primer_bind     complement(3232..3249)
                     /label=bghrev
     polyA_signal    3238..3462
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3508..3936
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3950..4279
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4130..4265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4327..4374
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4393..4767
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     complement(4599..4617)
                     /label=testtfprabcmvrev
     primer_bind     complement(4606..4617)
                     /label=mcherry3pletm1_rev
     primer_bind     complement(4606..4617)
                     /label=mcherry3pletm1_reva
     polyA_signal    4897..5018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4897..4914)
                     /label=EGFP-C-Rev
     primer_bind     complement(5067..5083)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5091..5107
                     /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(join(5115..5121,5122..5139,5140..5145))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5160..5181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5469..6054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6225..7016,7017..7085))
                     /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(7086..7190)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcag ctcagctttc tagaccgcgc
      721 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tccctatcag
      781 tgatagagat ctccctatca gtgatagaga tcgtcgacga gctcgtttag tgaaccgtca
      841 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
      901 cagcctccgg actctagcgt ttaaacttaa gcttggtacc gagctcggat ccatgtctac
      961 caggtctgtg tcctcgtcct cctaccgcag gatgttcggt ggctccggca catcgagccg
     1021 gcccagctcc aaccggagct atgtgaccac gtccacacgc acctacagtc tgggcagcgc
     1081 actgcgcccc agcactagcc gcagcctcta ttcctcatcc cccggtggcg cctatgtgac
     1141 ccggtcctcg gcagtgcgcc tgcggagcag cgtgccgggc gtgcggctgc ttcaagactc
     1201 ggtggacttc tcgctggccg acgccatcaa cactgagttc aagaacaccc gcaccaacga
     1261 gaaggtagaa ctgcaggagc tgaatgaccg ctttgccaac tacatcgaca aggtgcgctt
     1321 cctcgagcag cagaacaaaa tcctgctggc tgagctcgag cagctcaagg gccagggcaa
     1381 gtcgcgcctg ggcgacctgt acgaggagga gatgcgggag ctgcgccggc aggtggatca
     1441 gctcaccaac gacaaggccc gtgtcgaggt ggagcgggac aacctggccg aggacatcat
     1501 gcggctgcga gagaaattgc aggaggagat gctccagaga gaggaagccg aaagcaccct
     1561 gcagtcattc agacaggatg ttgacaatgc ttctctggca cgtcttgacc ttgaacggaa
     1621 agtggaatcc ttgcaggaag aaattgcctt tttgaagaaa ctgcacgatg aagagatcca
     1681 ggagctgcag gcccagattc aggaacagca tgtccagatc gatgtggacg tttccaagcc
     1741 tgacctcact gctgccctgc gtgatgtgcg ccagcagtat gaaagcgtgg ctgccaagaa
     1801 cctccaggag gccgaggaat ggtacaagtc caagtttgct gacctctctg aggctgccaa
     1861 ccggaacaac gatgccctgc gccaggccaa gcaggagtca aacgagtacc ggagacaggt
     1921 gcagtcactc acctgtgaag tggatgccct taaaggcact aacgagtccc tggagcgcca
     1981 gatgcgtgag atggaagaga attttgccct tgaagctgct aactaccagg acactattgg
     2041 ccgcctgcag gatgagatcc aaaacatgaa ggaagagatg gctcgtcacc ttcgtgaata
     2101 ccaagatctg ctcaatgtta agatggccct ggacattgag atcgccacct acaggaagct
     2161 gctggaaggc gaggagagca ggatttctct gcctctgcca accttttctt ccctgaacct
     2221 gagagaaact aacctggagt cacttcctct ggttgacacc cactcaaaaa gaacactcct
     2281 gattaagacg gttgagacca gagatggaca ggtgatcaat gagacttctc agcatcacga
     2341 tgaccttgaa gaattcggtg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     2401 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     2461 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     2521 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     2581 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     2641 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     2701 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     2761 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     2821 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     2881 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     2941 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     3001 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     3061 caagggtggt ggtggcgcgg ccgcagatcg atcgatcacg cgtgccacca tgactagttt
     3121 tgtacaaagc ggtccgtacc catacgatgt tccagattac gctaccgcgg taaacatatg
     3181 gatcgatcat aatactcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     3241 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3301 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3361 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3421 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3481 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3541 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3601 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3661 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3721 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3781 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3841 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3901 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3961 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4081 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4141 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4201 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4261 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4321 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     4381 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     4441 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     4501 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     4561 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     4621 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     4681 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     4741 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
     4801 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4861 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4921 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4981 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5041 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5101 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5161 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5221 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5281 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5341 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5401 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5461 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5521 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5581 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5641 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5701 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5761 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5821 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5881 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5941 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6001 agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6061 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6121 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6181 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6241 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6301 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6361 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6421 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6481 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6541 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6601 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6661 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6721 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6781 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6841 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6901 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6961 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7021 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7081 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7141 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7201 cgaaaagtgc cacctgacgt c
//
