LOCUS       pNTVimentin-mche        7251 bp    DNA     circular SYN 06-NOV-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7251)
  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..7251
                     /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"
     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"
     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    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"
     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"
     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"
     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..3164
                     /label=suntagv4for
     primer_bind     3136..3161
                     /label=testv4bspE1for
     misc_feature    3137..3193
                     /label=suntagv4-1
     misc_feature    3137..3193
                     /label=suntagv4-1
     primer_bind     complement(3169..3210)
                     /label=suntagv4rev
     primer_bind     complement(3169..3194)
                     /label=testv4hind3rev
     primer_bind     complement(3262..3279)
                     /label=BGH-rev
     polyA_signal    3268..3492
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3538..3966
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3980..4309
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4160..4295
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4357..4404
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4423..4797
                     /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(4629..4647)
                     /label=testtfprabcmvrev
     primer_bind     complement(4636..4647)
                     /label=mcherry3pletm1_rev
     primer_bind     complement(4636..4647)
                     /label=mcherry3pletm1_reva
     polyA_signal    4927..5048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4927..4944)
                     /label=EGFP-C-Rev
     primer_bind     complement(5097..5113)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5121..5137
                     /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(5145..5151,5152..5169,5170..5175))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5190..5211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5499..6084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6255..7046,7047..7115))
                     /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(7116..7220)
                     /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 ggtccggaag aacttttgag caagaattat catcttgaga acgaagtggc
     3181 tcgtcttaag aaaaccgcgg taaacatatg gatcgatcat aatactcgag tctagagggc
     3241 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3301 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3361 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3421 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3481 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3541 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3601 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3661 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3721 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3781 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3841 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3901 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3961 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4021 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4081 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4141 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4201 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4261 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4321 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
     4381 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
     4441 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     4501 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     4561 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     4621 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     4681 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     4741 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
     4801 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4861 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4921 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4981 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5041 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5101 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5161 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5221 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5281 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5341 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5401 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5461 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5521 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5581 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5641 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5701 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5761 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5821 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5881 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5941 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6001 tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt
     6061 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6121 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6181 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6241 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6301 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6361 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6421 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6481 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6541 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6601 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6661 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6721 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6781 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6841 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6901 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6961 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7021 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7081 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7141 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7201 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
