LOCUS       Exported                9764 bp DNA     circular SYN 27-SEP-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pDSK-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9764)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 27, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1079..1095
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             1314..1336
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1344..2054
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2103..2119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2127..2143
                     /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(2151..2181)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2196..2217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3719..4558)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(5069..6040)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     oriT            7279..7366
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      7708..8102
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     CDS             complement(8916..9731)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
       61 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
      121 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
      181 gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
      241 agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca
      301 ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg
      361 atgatggggc gattcaggcc tggtatgagt cagcaacacc ttcttcacga ggcagacctc
      421 agcgctattc tgaccttgcc atcacgactg tgctggtcat taaacgcgta ttcaggctga
      481 ccctgcgcgc tgcgcagggc tttattgatt ccatttttac actgatgaat gttccgttgc
      541 gctgcccgga ttacagctgc gtttcctgtt tggggtcgtt tgcgggaagg ggcggaatcc
      601 tacgctaagg ctttggccag cgatattctc cggtgagatt gatgtgttcc caggggatag
      661 gagaagtcgc ttgatatcta gtatgacgtc tgtcgcacct gcttgatcgc ggccgcgata
      721 gctagatcgc gttgctcctc ttctccatcc gcgttccaag ctgcggaaag gcacccataa
      781 gcgtacgcct ggtcgagcag gcgacgcgga tcgacgtcca gcgcacgaga gaatgcgtcc
      841 gccatctgtg caatgcgtct aggatcgaga caaaggtcgt ctctgtcagc cggatcgtag
      901 aacatattgg cggcgccaaa gcccacttca ccgaccagac attcgccatt caggctgcgc
      961 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     1021 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     1081 aaaacgacgg ccagtgaatt cgagctcggt acccggggat ctagaggatc tcaatgaata
     1141 ttggttgaca cgggcgtata agacatgtta tactgttgaa taacaagctt tcaattctct
     1201 attttgagaa gaattaatgt gcttgggagt ccctgatgat taaattaaat aaaccaagat
     1261 tttaccatga ctgcaatttt agagagacgg gggatcgatc ctctagaaat aattttgttt
     1321 aactttaaga aggagatata catatggtga gcaagggcga ggaggataac atggccatca
     1381 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     1441 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     1501 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     1561 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1621 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1681 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1741 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1801 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1861 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1921 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1981 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     2041 agctgtacaa gtaaggatcc tctagagtcg acctgcaggc atgcaagctt ggcgtaatca
     2101 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2161 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2221 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2281 atcggccaac gcgcggggag aggcggtttg cgtattgggc cacgcttccc gaagggagaa
     2341 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2401 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2461 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2521 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2581 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2641 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaagaaga gcatacatct
     2701 ggaagcaaag ccaggaaagc ggcctatgga gctgtgcggc agcgctcagt aggcaatttt
     2761 tcaaaatatt gttaagcctt ttctgagcat ggtatttttc atggtattac caattagcag
     2821 gaaaataagc cattgaatat aaaagataaa aatgtcttgt ttacaataga gtgggggggg
     2881 tcagcctgcc gccttgggcc gggtgatgtc gtacttgccc gccgcgaact cggttaccgt
     2941 ccagcccagc gcgaccagct ccggcaacgc ctcgcgcacc cgctggcggc gcttgcgcat
     3001 ggtcgaacca ctggcctctg acggccagac atagccgcac aaggtatcta tggaagcctt
     3061 gccggttttg ccggggtcga tccagccaca cagccgctgg tgcagcaggc gggcggtttc
     3121 gctgtccagc gcccgcacct cgtccatgct gatgcgcaca tgctggccgc cacccatgac
     3181 ggcctgcgcg atcaaggggt tcagggccac gtacaggcgc ccgtccgcct cgtcgctggc
     3241 gtactccgac agcagccgaa acccctgccg cttgcggcca ttctgggcga tgatggatac
     3301 cttccaaagg cgctcgatgc agtcctgtat gtgcttgagc gccccaccac tatcgacctc
     3361 tgccccgatt tcctttgcca gcgcccgata gctacctttg accaccatgg catcagcggt
     3421 gacggcctcc cacttgggtt ccaggaacag ccggagctgc cgtccgcctt cggtcttggg
     3481 ttccgggcca agcactaggc cattaggccc agccatggcc accagccctt gcaggatgcg
     3541 cagatcatca gcgcccagcg gctccgggcc gctgaactcg atccgcttgc cgtcgccgta
     3601 gtcatacgtc acgtccagct tgctgcgctt gcgctcgccc cgcttgaggg cacggaacag
     3661 gccgggggcc agacagtgcg ccgggtcgtg ccggacgtgg ctgaggctgt gcttgttctt
     3721 aggcttcacc acggggcacc cccttgctct tgcgctgcct ctccagcacg gcgggcttga
     3781 gcaccccgcc gtcatgccgc ctgaaccacc gatcagcgaa cggtgcgcca tagttggcct
     3841 tgctcacacc gaagcggacg aagaaccggc gctggtcgtc gtccacaccc cattcctcgg
     3901 cctcggcgct ggtcatgctc gacaggtagg actgccagcg gatgttatcg accagtaccg
     3961 agctgccccg gctggcctgc tgctggtcgc ctgcgcccat catggccgcg cccttgctgg
     4021 catggtgcag gaacacgata gagcacccgg tatcggcggc gatggcctcc atgcgaccga
     4081 tgacctgggc catggggccg ctggcgtttt cttcctcgat gtggaaccgg cgcagcgtgt
     4141 ccagcaccat caggcggcgg ccctcggcgg cgcgcttgag gccgtcgaac cactccgggg
     4201 ccatgatgtt gggcaggctg ccgatcagcg gctggatcag caggccgtca gccacggctt
     4261 gccgttcctc ggcgctgagg tgcgccccaa gggcgtgcag gcggtgatga atggcggtgg
     4321 gcgggtcttc ggcgggcagg tagatcaccg ggccggtggg cagttcgccc acctccagca
     4381 gatccggccc gcctgcaatc tgtgcggcca gttgcagggc cagcatggat ttaccggcac
     4441 caccgggcga caccagcgcc ccgaccgtac cggccaccat gttgggcaaa acgtagtcca
     4501 gcggtggcgg cgctgctgcg aacgcctcca gaatattgat aggcttatgg gtagccattg
     4561 attgcctcct ttgcaggcag ttggtggtta ggcgctggcg gggtcactac ccccgccctg
     4621 cgccgctctg agttcttcca ggcactcgcg cagcgcctcg tattcgtcgt cggtcagcca
     4681 gaacttgcgc tgacgcatcc ctttggcctt catgcgctcg gcatatcgcg cttggcgtac
     4741 agcgtcaggg ctggccagca ggtcgccggt ctgcttgtcc ttttggtctt tcatatcagt
     4801 caccgagaaa cttgccgggg ccgaaaggct tgtcttcgcg gaacaaggac aaggtgcagc
     4861 cgtcaaggtt aaggctggcc atatcagcga ctgaaaagcg gccagcctcg gccttgtttg
     4921 acgtataacc aaagccaccg ggcaaccaat agcccttgtc acttttgatc aggtagaccg
     4981 accctgaagc gcttttttcg tattccataa aacccccttc tgtgcgtgag tactcatagt
     5041 ataacaggcg tgagtaccaa cgcaagcact acatgctgaa atctggcccg cccctgtcca
     5101 tgcctcgctg gcggggtgcc ggtgcccgtg ccagctcggc ccgcgcaagc tggacgctgg
     5161 gcagacccat gaccttgctg acggtgcgct cgatgtaatc cgcttcgtgg ccgggcttgc
     5221 gctctgccag cgctgggctg gcctcggcca tggccttgcc gatttcctcg gcactgcggc
     5281 cccggctggc cagcttctgc gcggcgataa agtcgcactt gctgaggtca tcaccgaagc
     5341 gcttgaccag cccggccatc tcgctgcggt actcgtccag cgccgtgcgc cggtggcggc
     5401 taagctgccg ctcgggcagt tcgaggctgg ccagcctgcg ggccttctcc tgctgccgct
     5461 gggcctgctc gatctgctgg ccagcctgct gcaccagcgc cgggccagcg gtggcggtct
     5521 tgcccttgga ttcacgcagc agcacccacg gctgataacc ggcgcgggtg gtgtgcttgt
     5581 ccttgcggtt ggtgaagccc gccaagcggc catagtggcg gctgtcggcg ctggccgggt
     5641 cggcgtcgta ctcgctggcc agcgtccggg caatctgccc ccgaagttca ccgcctgcgg
     5701 cgtcggccac cttgacccat gcctgatagt tcttcgggct ggtttccact accagggcag
     5761 gctcccggcc ctcggctttc atgtcatcca ggtcaaactc gctgaggtcg tccaccagca
     5821 ccagaccatg ccgctcctgc tcggcgggcc tgatatacac gtcattgccc tgggcattca
     5881 tccgcttgag ccatggcgtg ttctggagca cttcggcggc tgaccattcc cggttcatca
     5941 tctggccggt ggtggcgtcc ctgacgccga tatcgaagcg ctcacagccc atggccttga
     6001 gctgtcggcc tatggcctgc aaagtcctgt cgttcttcat cgggccacca agcgcagcca
     6061 gatcgagccg tcctcggttg tcagtggcgt caggtcgagc aagagcaacg atgcgatcag
     6121 cagcaccacc gtaggcatca tggaagccag catcacggtt agccatagct tccagtgcca
     6181 cccccgcgac gcgctccggg cgctctgcgc ggcgctgctc acctcggcgg ctacctcccg
     6241 caactctttg gccagctcca cccatgccgc ccctgtctgg cgctgggctt tcagccactc
     6301 cgccgcctgc gcctcgctgg cctgctgggt ctggctcatg acctgccggg cttcgtcggc
     6361 cagtgtcgcc atgctctggg ccagcggttc gatctgctcc gctaactcgt tgatgcctct
     6421 ggatttcttc actctgtcga ttgcgttcat ggtctattgc ctcccggtat tcctgtaagt
     6481 cgatgatctg ggcgttggcg gtgtcgatgt tcagggccac gtctgcccgg tcggtgcgga
     6541 tgccccggcc ttccatctcc accacgttcg gccccaggtg aacaccgggc aggcgctcga
     6601 tgccctgcgc ctcaagtgtt ctgtggtcaa tgcgggcgtc gtggccagcc cgctctaatg
     6661 cccggttggc atggtcggcc catgcctcgc gggtctgctc aagccatgcc ttgggcttga
     6721 gcgcttcggt cttctgtgcc ccgcccttct ccggggtctt gccgttgtac cgcttgaacc
     6781 actgagcggc gggccgctcg atgccgtcat tgatccgctc ggagatcatc aggtggcagt
     6841 gcgggttctc gccgccaccg gcatggatgg ccagcgtata cggcaggcgc tcggcaccgg
     6901 tcaggtgctg ggcgaactcg gacgccagcg ccttctgctg gtcgagggtc agctcgaccg
     6961 gcagggcaaa ttcgacctcc ttgaacagcc gcccattggc gcgttcatac aggtcggcag
     7021 catcccagta gtcggcgggc cgctcgacga actccggcat gtgcccggat tcggcgtgca
     7081 agacttcatc catgtcgcgg gcatacttgc cttcgcgctg gatgtagtcg gccttggccc
     7141 tggccgattg gccgcccgac ctgctgccgg ttttcgccgt aaggtgataa atcgccatgc
     7201 tgcctcgctg ttgcttttgc ttttcggctc catgcaatgg ccctcggaga gcgcaccgcc
     7261 cgaagggtgg ccgttaggcc agtttctcga agagaaaccg gtaagtgcgc cctcccctac
     7321 aaagtagggt cgggattgcc gccgctgtgc ctccatgata gcctacgaga cagcacatta
     7381 acaatggggt gtcaagatgg ttaaggggag caacaaggcg gcggatcggc tggccaagct
     7441 cgaagaacaa cgagcgcgaa tcaatgccga aattcagcgg gtgcgggcaa gggaacagca
     7501 gcaagagcgc aagaacgaaa caaggcgcaa ggtgctggtg ggggccatga ttttggccaa
     7561 ggtgaacagc agcgagtggc cggaggatcg gctcatggcg gcaatggatg cgtaccttga
     7621 acgcgaccac gaccgcgcct tgttcggtct gccgccacgc cagaaggatg agccgggctg
     7681 aatgatcgac cgagacaggc cctgcggggc tgcacacgcg cccccaccct tcgggtaggg
     7741 ggaaaggccg ctaaagcggc taaaagcgct ccagcgtatt tctgcggggt ttggtgtggg
     7801 gtttagcggg ctttgcccgc ctttccccct gccgcgcagc ggtggggcgg tgtgtagcct
     7861 agcgcagcga atagaccagc tatccggcct ctggccgggc atattgggca agggcagcag
     7921 cgccccacaa gggcgctgat aaccgcgcct agtggattat tcttagataa tcatggatgg
     7981 atttttccaa caccccgcca gcccccgccc ctgctgggtt tgcaggtttg ggggcgtgac
     8041 agttattgca ggggttcgtg acagttattg caggggggcg tgacagttat tgcaggggtt
     8101 cgtgacagtt agtacgggag tgacgggcac tggctggcaa tgtctagcaa cggcaggcat
     8161 ttcggctgag ggtaaaagaa ctttccgcta agcgatagac tgtatgtaaa cacagtattg
     8221 caaggacgcg gaacatgcct catgtggcgg ccaggacggc cagccgggat cgggatactg
     8281 gtcgttacca gagccaccga cccgagcaaa cccttctcta tcagatcgtt gacgagtatt
     8341 acccggcatt cgctgcgctt atggcagagc agggaaagga attgccgggc tatgtgcaac
     8401 gggaatttga agaatttctc caatgcgggc ggctggagca tggctttcta cgggttcgct
     8461 gcgagtcttg ccacgccgag cacctggtcg ctttcagctg taatccgggc agcgcaacgg
     8521 aacattcatc agtgtaaaaa tggaatcaat aaagccctgc gcagcgcgca gggtcagcct
     8581 gaatacgcgt ttaatgacca gcacagtcgt gatggcaagg tcagaatagc gctgaggtct
     8641 gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga atcgccccat catccagcca
     8701 gaaagtgagg gagccacggt tgatgagagc tttgttgtag gtggaccagt tggtgatttt
     8761 gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt
     8821 caactcagca aaagttcgat ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg
     8881 ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa
     8941 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     9001 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     9061 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     9121 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     9181 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     9241 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     9301 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     9361 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     9421 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     9481 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     9541 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     9601 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     9661 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
     9721 atatggctca tagcacccct tgtattactg tttatgtaag caga
//
