LOCUS       pH2B_mcherry_HA         6214 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 6214)
  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..6214
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     720..740
                     /label=CMV-for
     primer_bind     720..740
                     /label=CMVfor
     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"
     gene            963..1340
                     /label=H2B(HIST1H2BJ)
     primer_bind     1345..1370
                     /label=mTFPmito_for
     CDS             1353..2057
                     /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     1353..1367
                     /label=mcherry5pletm1_for
     primer_bind     1991..2006
                     /label=mcherry for
     primer_bind     complement(2029..2057)
                     /label=mtfprab5f.REV
     primer_bind     complement(2039..2081)
                     /label=mTFPmito_rev
     primer_bind     complement(2043..2077)
                     /label=mcherry3pletm1_reva
     primer_bind     complement(2043..2077)
                     /label=mcherry3pletm1_rev
     primer_bind     2118..2146
                     /label=HAformito_for
     primer_bind     2129..2156
                     /label=testhabspe1for
     primer_bind     complement(2130..2157)
                     /label=testhahind3rev
     CDS             2130..2156
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2142..2173)
                     /label=HAformito_rev
     primer_bind     complement(2225..2242)
                     /label=bghrev
     primer_bind     complement(2225..2242)
                     /label=BGH-rev
     polyA_signal    2231..2455
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2501..2929
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2943..3272
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3123..3258
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3320..3367
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3386..3760
                     /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(3599..3610)
                     /label=mcherry3pletm1_reva
     primer_bind     complement(3599..3610)
                     /label=mcherry3pletm1_rev
     polyA_signal    3890..4011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4060..4076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4084..4100
                     /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(4108..4114,4115..4132,4133..4138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4153..4174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4462..5047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5218..6009,6010..6078))
                     /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(6079..6183)
                     /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 cgtaccgcca
      961 ccatgccaga gccagcgaag tctgctcccg ccccgaaaaa gggctccaag aaggcggtga
     1021 ctaaggcgca gaagaaaggc ggcaagaagc gcaagcgcag ccgcaaggag agctattcca
     1081 tctatgtgta caaggttctg aagcaggtcc accctgacac cggcatttcg tccaaggcca
     1141 tgggcatcat gaattcgttt gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc
     1201 gcctggcgca ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc
     1261 gcctgctgct gcctggggag ttggccaagc acgccgtgtc cgagggtact aaggccatca
     1321 ccaagtacac cagcgctaag gatcaattcg gtgtgagcaa gggcgaggag gataacatgg
     1381 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     1441 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     1501 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     1561 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     1621 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     1681 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     1741 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     1801 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     1861 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     1921 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     1981 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     2041 tggacgagct gtacaagggt ggtggtggcg cggccgcaga tcgatcgatc acgcgtgcca
     2101 ccatgactag ttttgtacaa agcggtccgt acccatacga tgttccagat tacgctaccg
     2161 cggtaaacat atggatcgat cataatactc gagtctagag ggcccgttta aacccgctga
     2221 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2281 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2341 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2401 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2461 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2521 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2581 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2641 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2701 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2761 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2821 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2881 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2941 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3001 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3061 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3121 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3181 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3241 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3301 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     3361 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
     3421 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     3481 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     3541 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     3601 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     3661 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     3721 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
     3781 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3841 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3901 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3961 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4021 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4081 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4141 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4201 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4261 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4321 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4381 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4441 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4501 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4561 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4621 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4681 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4741 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4801 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4861 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4921 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4981 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5041 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5101 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5161 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5221 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5281 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5341 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5401 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5461 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5521 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5581 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5641 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5701 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5761 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5821 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5881 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5941 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6001 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6061 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6121 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6181 gcgcacattt ccccgaaaag tgccacctga cgtc
//
