LOCUS       mitomcherryHA           6024 bp    DNA     circular SYN 23-MAY-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6024)
  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..6024
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     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"
     misc_feature    959..1135
                     /label=CISD1 (MitoNEET)
     primer_bind     1144..1177
                     /label=mcherry5pletm1_for
     primer_bind     1153..1180
                     /label=mTFPmito_for
     CDS             1163..1867
                     /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     1801..1816
                     /label=mcherry for
     primer_bind     complement(1839..1867)
                     /label=mtfprab5f.REV
     primer_bind     complement(1849..1891)
                     /label=mTFPmito_rev
     primer_bind     complement(1853..1887)
                     /label=mcherry3pletm1_rev
     primer_bind     complement(1853..1887)
                     /label=mcherry3pletm1_reva
     primer_bind     1928..1956
                     /label=HAformito_for
     primer_bind     1939..1966
                     /label=testhabspe1for
     primer_bind     complement(1940..1967)
                     /label=testhahind3rev
     CDS             1940..1966
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1952..1983)
                     /label=HAformito_rev
     primer_bind     complement(2035..2052)
                     /label=BGH-rev
     primer_bind     complement(2035..2052)
                     /label=bghrev
     polyA_signal    2041..2265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2311..2739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2753..3082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2933..3068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3130..3177
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3196..3570
                     /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(3409..3420)
                     /label=mcherry3pletm1_rev
     primer_bind     complement(3409..3420)
                     /label=mcherry3pletm1_reva
     polyA_signal    3700..3821
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3870..3886)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3894..3910
                     /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(3918..3924,3925..3942,3943..3948))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3963..3984
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4272..4857)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5028..5819,5820..5888))
                     /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(5889..5993)
                     /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 ccgccaccat
      961 gagtctgact tccagttcca gcgtacgagt tgaatggatc gcagcagtta ccattgctgc
     1021 tgggacagct gcaattggtt atctagctta caaaagattt tatgttaaag atcatcgaaa
     1081 taaagctatg ataaaccttc acatccagaa agacaacccc aagatagtac atgctggaag
     1141 cttaccggtc gccgaattcg gtgtgagcaa gggcgaggag gataacatgg ccatcatcaa
     1201 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
     1261 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     1321 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     1381 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     1441 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     1501 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     1561 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     1621 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     1681 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     1741 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     1801 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     1861 gtacaagggt ggtggtggcg cggccgcaga tcgatcgatc acgcgtgcca ccatgactag
     1921 ttttgtacaa agcggtccgt acccatacga tgttccagat tacgctaccg cggtaaacat
     1981 atggatcgat cataatactc gagtctagag ggcccgttta aacccgctga tcagcctcga
     2041 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2101 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2161 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2221 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     2281 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2341 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2401 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2461 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2521 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2581 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2641 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2701 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     2761 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2821 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2881 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2941 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3001 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3061 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3121 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     3181 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     3241 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     3301 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     3361 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     3421 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     3481 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     3541 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
     3601 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3661 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     3721 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3781 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     3841 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3901 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3961 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4021 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4081 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4141 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4201 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4261 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4321 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4381 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4441 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4501 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4561 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4621 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4681 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4741 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4801 ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4861 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4921 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4981 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5041 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5101 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5161 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5221 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5281 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5341 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5401 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5461 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5521 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5581 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5641 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5701 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5761 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5821 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5881 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5941 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6001 ccccgaaaag tgccacctga cgtc
//
