LOCUS       mitomcherrysunta        6054 bp    DNA     circular SYN 18-MAY-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6054)
  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..6054
                     /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"
     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     1163..1180
                     /label=testtfprab5cmvfor
     primer_bind     1801..1816
                     /label=mcherry for
     primer_bind     complement(1839..1867)
                     /label=mtfprab5f.REV
     primer_bind     complement(1846..1866)
                     /label=testtfprabcmvrev
     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..1967
                     /label=suntagv4for
     primer_bind     1939..1964
                     /label=testv4bspE1for
     misc_feature    1940..1996
                     /label=suntagv4-1
     primer_bind     complement(1972..2013)
                     /label=suntagv4rev
     primer_bind     complement(1972..1997)
                     /label=testv4hind3rev
     primer_bind     complement(2065..2082)
                     /label=BGH-rev
     polyA_signal    2071..2295
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2341..2769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2783..3112
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2963..3098
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3160..3207
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3226..3600
                     /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(3432..3450)
                     /label=testtfprabcmvrev
     primer_bind     complement(3439..3450)
                     /label=mcherry3pletm1_rev
     primer_bind     complement(3439..3450)
                     /label=mcherry3pletm1_reva
     polyA_signal    3730..3851
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3730..3747)
                     /label=EGFP-C-Rev
     primer_bind     complement(3900..3916)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3924..3940
                     /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(3948..3954,3955..3972,3973..3978))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3993..4014
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4302..4887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5058..5849,5850..5918))
                     /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(5919..6023)
                     /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 agcggtccgg aagaactttt gagcaagaat tatcatcttg agaacgaagt
     1981 ggctcgtctt aagaaaaccg cggtaaacat atggatcgat cataatactc gagtctagag
     2041 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2101 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2161 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2221 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2281 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2341 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2401 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2461 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2521 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2581 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2641 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2701 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2761 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2821 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2881 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2941 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3001 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3061 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3121 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     3181 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
     3241 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     3301 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     3361 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     3421 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     3481 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     3541 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     3601 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3661 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3721 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3781 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3841 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3901 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3961 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4021 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4081 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4141 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4201 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4261 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4321 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4381 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4441 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4501 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4561 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4621 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4681 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4741 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4801 agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag
     4861 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4921 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4981 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5041 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5101 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5161 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5221 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5281 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5341 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5401 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5461 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5521 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5581 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5641 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5701 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5761 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5821 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5881 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5941 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6001 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
