LOCUS       sequence-7754-ex        8804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8804)
  AUTHORS   Matsuda T, Cepko CL
  TITLE     Controlled expression of transgenes introduced by in vivo 
            electroporation.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 Jan 16;104(3):1027-32. doi: 
            10.1073/pnas.0610155104
  PUBMED    17209010
REFERENCE   2  (bases 1 to 8804)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1073/pnas"; 
            journalName: "Proc Natl Acad Sci U S A"; date: "2007-01-16- 16"; 
            volume: "104"; issue: "3"; pages: "1027-32"
COMMENT     Sequence Label: pCrx-DsRed
FEATURES             Location/Qualifiers
     source          1..8804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      5060..5069
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5066..5743
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(5246..5266)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     5658..5681
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(5828..5847)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5893..5948
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5947..5966)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6307..6323)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6307..6323)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6320..6342)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6331..6347
                     /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(6355..6361,6362..6379,6380..6385))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6400..6421
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6480..6675
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6526..6661
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6681..6815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6718..6737)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6772..6791
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6883..6900)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7054..7642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7134..7153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7813..8604,8605..8673))
                     /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"
     primer_bind     8436..8455
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8674..8778)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agtgatgggc catgtgaggg atagaatgga aacctaatat
       61 agtacaagct tcctaaaata catacacata tgaaggtgat ctaaatgaaa ctgccaaata
      121 atgggggggg ggcagagctt caggtggcca tctcttgtca ccaaacgaag cttccagcac
      181 caggactggg ttatacttaa ttgagttgtt ggtcaaaggg gtcccttggg aatccccaag
      241 ccacccaggc tgttgacaag actacaggtt gctcttcaca aactgatggc aaggccccac
      301 tgctgaagac aacacctacc caactcatta aacacggaga agtcaaagct ggtgcctaca
      361 cagaaccttc atgtctatgt tctagtgtct ctggtacagg aagatactct gcaaactacc
      421 aaaagatata tgtaaatact aacccagcca cagacctttt gatctacagt ggtgtcctgc
      481 ctgcaagata ggctagggca gtgccagcat gaagtttgtg ggagtggcca accactatct
      541 gacttaaagc ttactctgtg agatggaact catattgaca ctgcttatat gaccaagtac
      601 ctggactaga taaccaagga acctagggta aagccaaatg ctactattaa aaagaaaaag
      661 caggaggctg gctcagtggt taagagcact gactgctctt ccgaaggtcc taagttcaaa
      721 tcccagcaag cacacagtgg ttcacaacca tctgtaatga gatctgacac ccttttctgg
      781 tatgtctgaa gacagctata gtgtacttac atataataaa taaatctttg ggccagagtg
      841 gcagggctgg agtcagcaga ggtcctgagt tcaattccca gcaaccacat gatggctcac
      901 aaccatctac acagctacgg tgtactcata tacataaaat aagtaaataa aaaaattttt
      961 tttcgagaca gggtttttct gtatagccct ggctgtctcg gaactcactt tgtagaccag
     1021 gctggcctcg aactcagaaa tccacctgcc tctgcctccc gagtgctggg attaaaggtg
     1081 tgcgtcacta cccccggctc aataaatctt aaaaaaagaa aaaaagcaat aaaatgactc
     1141 ctaatggctt tcttttactc atagatcagt gccttgctca gccatcatca gagaaggttc
     1201 ctcctgcagc agatgggaag agatacagag atccacggct agacatatgc agagagctga
     1261 agttctggag aactcagctc taaaggggtg tctccattat atccatacct caactcctgc
     1321 tgtgcaggga atcctgcaaa agaggaggaa gaaagagtgt atgagtcaga agggatggag
     1381 gccaccaagg aaacaaggtt ctctaaactg acatattcaa agctcatgtg aactcacaga
     1441 tcccgaggca gcatgcacag ggcctgtgtg gctctgcacc cgggcttctg cctatttgtt
     1501 aattattctc gttaaattct tttgtgggat cctgagtgtg agaacagatc ttatcttgcg
     1561 ccttctcttg gttttttgaa cttctctcaa ctgtgactct tagtttttac tttattttat
     1621 tattatattt cattgaatta ttatccctta taagcctgtt tgttttctag tgagacagaa
     1681 agggagtggc tccagatgag aggggaaagt ggggtgtaac tgggaggagt agagtgtggg
     1741 gaaaccgaaa tcagatatta tgtaagaaaa agaaatctat ttttactaac gagaaaaaaa
     1801 gagtaattcc ttgtcagatg tgatgagcac acacctgtaa tcccagttct gacgagacag
     1861 gtagatctct gtaaattcaa ggctagcctg gtctacatag gtacaaacag ctgagccagt
     1921 gctacaaaag tgagacctgt ctcaaaaaaa ctacctcaca acacaaacaa aaagggggat
     1981 tgcctggagg tctgatcctt aacacatcaa aacagaaaat gacaacaaac aaacaaacaa
     2041 taaacagcaa caaacccaaa accctaccta agcaacaaag ccaaaagaat gtctttccag
     2101 aaatctctga ggtcctgggg ctatctgtgg aggtctgtcc ttgggtgggg caaggcagga
     2161 attatagcaa ggacatctgc tctcagcagc caccagctcc tcctccatgc ttcccagtga
     2221 caccatagga gctcccagaa ccaccaggga ggtttcctca tctattccag tcagtctcac
     2281 tcattctctc agccttcatc agtgtcccac tgaacacaag tgccaggcca gcaaggtggc
     2341 tcagcaggta aaaagtactt gccaccaagt atccccaggt ccccatggtg gaagaataga
     2401 accggcttct gcaggttgtc tttcgtctct tccatgtgaa catcctgctt tccacaaaca
     2461 aacaaacaaa caaacaaaca aacaaacaat gtagttaaac caattcagac atgtcatgaa
     2521 ttcataatta tggatgaaaa gacagtccct atagagaggg gacacatttc aattatttgg
     2581 gtagaggcat ctagagctag ggtgatcctg agggagagcc aagttaggta cccccccccc
     2641 cacacacaca cacaatgtag agattgaacc tagaaccttg ctcttgccag gccttttagt
     2701 cccagtcttt caagttttct aaaacaaaag caaaaacaaa aaaatctaaa caaggtgggg
     2761 cttgttgatg cacgccttta ttcccagcac tgggaggcag gggcaagtgg atctatgtgg
     2821 gtttgaggcc agcctggtct acagaacaag tataggacag ccatggctct ttacacagag
     2881 aaaccctatc tcgaaaaaac aaaacaaaac aacaacaatc ccaaacaaaa aacaaaatac
     2941 ggaccccctc aaaacaacaa caacaacaac aacaacaaca acaacaacaa caacaacaac
     3001 aacaaaaatc aaccctccaa aactaaacca gaaaacccgc tcgtcacatg cattctgtag
     3061 gactttaggc tagaaatcta aagatgaggc ccctggcctg aaggattgtt tcctaacccc
     3121 gtctgcatag tttagccttc ctactgggct gagcaaaaac gtgggcctga gcctcctgtc
     3181 aagcttcaga gagcgtcaga caactcctcc tttgtgtcat tctactaatg agatctgctc
     3241 tgattccctt tctggcccag aggccctaat ctctcctagc agggggctgg ggatagggga
     3301 cagaggggag gtcccctcct gacaagcagt gacccagtgg tgagcaggga ttagagtgtg
     3361 cgtgtggctg tgtgtgcctg tgagtgcacg gcccggtgtt tagagaagca gaaccaattt
     3421 gagtggggaa tgggactcga agtgatatta atacctagag agatgagggg agggacagag
     3481 ggggagggaa ccctgagggg gagagagaga tggcaggaag atgtgtatgc gtggagtagc
     3541 aagcgaacct tcatttcagc tcaaggattg caggaagcaa ctagaggctc tgagaccagg
     3601 ctgtagcctc aggcttaggt ggacaggcag aagggtctca aatccaggaa gggacccaca
     3661 gacagtggca gaacgacaaa ggagggagac acagagggag acaatgagag aatgcaaact
     3721 tgccttacaa agggaactag ggacagagtc agagacaagg tgaagggctg gagggatggc
     3781 tctgtggtta agagcactgg ggctctccag gaggaccagg gctccattct caatacttaa
     3841 tggtggctta taacctgtaa ttcccggtcc agggcatctg atgccctctt ctggtctctt
     3901 caggcatcag gcactcactg tgtgtgaaca cgtgggtgca ggcacaatat ccatgtatgt
     3961 ataatcaata aacaaatcta aacaagagag acacagagag aaagagtcag acatacagag
     4021 acactcagag ataaaaagac agagagaaag agagacaggg aggaaaagag ccagaaccag
     4081 agctgcagag agacagccag agacagagag atagcgggag acagagagta gaggtagagt
     4141 ccgacacaga tggagactaa cagagagaca gagagacagt gggagacaga gagtctaaca
     4201 gagacacagg cagagattga cagagacata gaaactcagg gagaggcata gagacaaaga
     4261 cacaaagaaa ctcagagagg cagggtctga aatagtgatg aagatagaga agcagagact
     4321 agaggcaatg gagaagatac agagcaatgg ggagacagag atatagagag ccctgaggaa
     4381 gtgagggaag aagggagaaa gacagagagg cacagagaaa gagagagaaa ctgaggcaga
     4441 aaaagggaga accatcaaga actacacatt tgaattagtt cactcagtgg ggtcacagat
     4501 ctctcttccc agttattcta gctgtgtcag aagacttaaa agttcaatac tagcctgggc
     4561 cacaaagaac aacccaggaa gtcctgtgcc tttcttttct gtctcccagg agcactgtac
     4621 acttctttct gtgcttttga agacatttct tttcagatag tagctgaagc tggaggaata
     4681 ccctttagac ccaaggaaag acttccctga gaagtggggg ctcctggttg caggcaggag
     4741 ttgggctttc cctcccttct tcatcaaccc taattgccaa gatgtcactg tgggggtggt
     4801 gggattaagc agacggtgcc cttcccccca gccaatgtca cctcctgttg cccagtgagt
     4861 cccccacctt ccctgggatt acccttccag atccaggcca caatagatga catcactccc
     4921 agcccaggct taaagtcgcc ccgtgtgagg ggacctattt cctctagcct ctgctgtctg
     4981 atgacaccat ccagtacctg aacatccagg agagtcccca tttctcatcg aattcccgcg
     5041 ggcccgggat ccaccggtcg ccaccatggc ctcctccgag aacgtcatca ccgagttcat
     5101 gcgcttcaag gtgcgcatgg agggcaccgt gaacggccac gagttcgaga tcgagggcga
     5161 gggcgagggc cgcccctacg agggccacaa caccgtgaag ctgaaggtga ccaagggcgg
     5221 ccccctgccc ttcgcctggg acatcctgtc cccccagttc cagtacggct ccaaggtgta
     5281 cgtgaagcac cccgccgaca tccccgacta caagaagctg tccttccccg agggcttcaa
     5341 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggcg accgtgaccc aggactcctc
     5401 cctgcaggac ggctgcttca tctacaaggt gaagttcatc ggcgtgaact tcccctccga
     5461 cggccccgtg atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc
     5521 ccgcgacggc gtgctgaagg gcgagaccca caaggccctg aagctgaagg acggcggcca
     5581 ctacctggtg gagttcaagt ccatctacat ggccaagaag cccgtgcagc tgcccggcta
     5641 ctactacgtg gacgccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     5701 gcagtacgag cgcaccgagg gccgccacca cctgttcctg tagcggccgc actcctcagg
     5761 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     5821 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     5881 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     5941 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     6001 tggtttagag tttggcaaca tatgccatat gctggctgcc atgaacaaag gtggctataa
     6061 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     6121 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     6181 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     6241 gtcatagctg tccctcttct cttatgaaga tccctcgacc tgcagcccaa gcttggcgta
     6301 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6361 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6421 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg
     6481 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     6541 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6601 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6661 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     6721 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6781 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     6841 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6901 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6961 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7021 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7081 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7141 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7201 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     7261 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7321 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7381 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7441 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7501 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7561 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7621 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7681 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7741 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7801 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7861 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7921 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7981 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8041 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8101 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8161 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8221 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8281 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8341 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8401 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8461 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8521 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8581 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8641 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8701 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8761 acaaataggg gttccgcgca catttccccg aaaagtgcca cctg
//
