LOCUS       psuntagV4_mcherr        6154 bp    DNA     circular SYN 15-JUN-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6154)
  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..6154
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     primer_bind     135..152
                     /label=pcdnafor
     primer_bind     223..263
                     /label=testcmvpromfor
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     726..745
                     /label=EGFP-N-for
     primer_bind     769..789
                     /label=CMVfor
     primer_bind     769..789
                     /label=CMV-for
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    841..859
                     /label=tet operator
     primer_bind     954..988
                     /label=testv4bspE1for
     primer_bind     963..991
                     /label=suntagv4for
     misc_feature    964..1020
                     /label=suntagv4-1
     primer_bind     complement(996..1025)
                     /label=testv4hind3rev
     primer_bind     complement(996..1021)
                     /label=suntagv4rev
     primer_bind     1025..1050
                     /label=mcherry5pletm1_for
     primer_bind     1031..1053
                     /label=testtfprab5cmvfor
     CDS             1033..1740
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFSEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1033..1053
                     /label=mTFPmito_for
     primer_bind     complement(1712..1740)
                     /label=mtfprab5f.REV
     primer_bind     complement(1712..1740)
                     /label=mtfprab5f.REVa
     primer_bind     complement(1719..1739)
                     /label=testtfprabcmvrev
     primer_bind     complement(1722..1741)
                     /label=mTFPmito_rev
     primer_bind     1746..1764
                     /label=h2bnterm_for
     misc_feature    1747..2127
                     /label=HIST1H2BJ
     primer_bind     complement(2105..2124)
                     /label=h2bnterm_rev
     primer_bind     complement(2165..2182)
                     /label=BGH-rev
     polyA_signal    2171..2395
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2441..2869
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2883..3212
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3063..3198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3260..3307
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3326..3700
                     /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(3532..3550)
                     /label=testtfprabcmvrev
     polyA_signal    3830..3951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3830..3847)
                     /label=EGFP-C-Rev
     primer_bind     complement(4000..4016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4024..4040
                     /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(4048..4054,4055..4072,4073..4078))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4093..4114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4402..4987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5158..5949,5950..6018))
                     /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(6019..6123)
                     /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 ttgacgtcaa tgggagtttg ttttggaacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 atggaagaac ttttgagcaa gaattatcat cttgagaacg aagtggctcg tcttaagaaa
     1021 aagctcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     1081 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     1141 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     1201 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     1261 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttctc cgagggcttc
     1321 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     1381 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1441 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1501 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1561 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1621 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1681 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1741 gatatcatgc cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg
     1801 gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat
     1861 tccatctatg tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag
     1921 gccatgggca tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct
     1981 tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc
     2041 gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc
     2101 atcaccaagt acaccagcgc taagtgactc gagtctagag ggcccgttta aacccgctga
     2161 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2221 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2281 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2341 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2401 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2461 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2521 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2581 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2641 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2701 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2761 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2821 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2881 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2941 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3061 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3121 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3181 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3241 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     3301 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
     3361 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     3421 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     3481 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     3541 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     3601 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     3661 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
     3721 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3781 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3841 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3901 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3961 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4021 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4081 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4141 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4201 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4261 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4321 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4381 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4441 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4501 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4561 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4621 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4681 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4741 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4801 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4861 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4921 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4981 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5041 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5101 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5161 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5221 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5281 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5341 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5401 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5461 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5521 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5581 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5641 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5701 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5761 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5821 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5881 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5941 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6001 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6061 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6121 gcgcacattt ccccgaaaag tgccacctga cgtc
//
