LOCUS       sequence-7753-ex        7005 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 7005)
  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 7005)
  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: pNrl-DsRed
FEATURES             Location/Qualifiers
     source          1..7005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      3261..3270
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3267..3944
                     /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(3447..3467)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3859..3882
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4029..4048)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4094..4149
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4148..4167)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4508..4524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4508..4524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4521..4543)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4532..4548
                     /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(4556..4562,4563..4580,4581..4586))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4601..4622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4681..4876
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4727..4862
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4882..5016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4919..4938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4973..4992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5084..5101)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5255..5843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5335..5354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6014..6805,6806..6874))
                     /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     6637..6656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6875..6979)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacttac atgtacatac ttcataatgg tctagaaggg cttattcaaa tttggcgttt
       61 actcagagtc ctcggggctc ttatccggga tgtggtgatg atgtgaggag aggcaactct
      121 caaactagga ccatcactcg gggcttgtgt atcagtagtg aggtccagca atggaaggga
      181 aatcccggga aaaaagtcta tgtgatccct aggagcaggt ctgagctctt tgtgcagaaa
      241 atttctggat acctggagta tgattctcag ggaccagggg tctgggtcag agaagggtgg
      301 ctgtctcaca tcatggggta cattcagcta tagaaagacc agcatgaacc aggcagtggt
      361 acctggcctg aaccaggact ggctcagctt gcttgagctt atttggcccg actcccttca
      421 ctccttagca gcttagctct cccacctctc tcctgagtcc tccagtaaac aagaggatca
      481 ggacagatat gacagctgcc tggtggggca agtgtctggt caggtcaaca gcaggcatga
      541 agggggctac tacactaaat gctgccaacc tttttccttt agggaagttc taggatctct
      601 ttcccggata tccactgtaa aacctcattc cactggcatg tatccctatt cctttcagca
      661 tccctattgc ccttttgtgg atggttcctg gccttacatt ttccgcttct tttaagaaaa
      721 taagcaacag gtcttggtga agagtaacat ttccattcga atagtgactc tatttcataa
      781 accaaacgaa acgaaacgaa acgaaacgaa acgaaacaaa acaaaacaaa acaaaacacc
      841 aagagacctg ggcatgggag agttgtgata tctgacctaa gccttctaat gtatgtagag
      901 gaagcttgga ttccaaatgt cctccaaagc tgggcatggt ggtgcgtgcc tttacgccct
      961 gtactaggaa ggcacaagca ggtagatcta tgggttcaag gtcaatctgg tctacaaaga
     1021 gagttctagg acggctaggg ctacacagag aaaccctgtt ttgaaaacca aaccaaacca
     1081 aaccaatgtc ctccaaaggc atttgagttc aaaacttgtc attattggaa ggtggtgaaa
     1141 ttttcaagag gtggggccct gtgggaagtc ttctggtctt tgggttttcc ctgaaggaag
     1201 caaaggccta acccctccct ggtcatcggt tggtaccctt gagggaagca ggagtcctat
     1261 ccacttccct tcccttttct tttttttttt ttttgtccca gtgataaggt gagtagcttt
     1321 cagttgtgtc ctcgccataa cacttgactc actacaggca atacatgatg ggaccaactg
     1381 ctcactggct aggctctcca aggcagcaaa ccaaagcaaa gcgctttccg attaagtcaa
     1441 ttacttcagg tatgtattgg agcaattatg ttagtgttag tcggtttttg ttattgtgac
     1501 aaagtgcccg agaaagatca aaggagggga gccttctttg ggctaatggc ttcagtttca
     1561 gctgatggtt gtgggtgtcc tattgctttg gtccaaaatc taacatcacg atggaaaatg
     1621 tatagtgaag aagtaaagag agaaaagaga gaggaagagg gggaagtgga ggtgggggag
     1681 aaggagggat ggagggacac agacaaaatg caccaatgaa ctaaggagaa gatacaccat
     1741 tcaaacgcat gttttttcta gttaggccaa ttcatactga cagccaattc agtatggatt
     1801 cttcagctgg ttagttatgg ataagtttaa cctctaataa tatctcagtg aaaactagct
     1861 tttaaaatat accggaacca tagcagtgat agaaaactaa tgtagggtgg acacacacac
     1921 acacacacac acacaaacac acactttgag actgggttga aggtaggcat ctgtggacca
     1981 gtgggtctca aacattagtg tgtttcagag ctgttggaag ggtttattaa acagcagctg
     2041 tctataacct acttactgag tttctggttc tgtgagtttg aggtaaagca aaagaatttg
     2101 tttcttcaag ttcccaaggg gtgatttctt aagactgtgg aagccattgt tccaacaggc
     2161 agggaatacc ctttaacttc tggagtttct tttatcacag cacctattta aacagcttca
     2221 ggcaggttta aacatctcct tggctaagtc ctactatccc tgcttcggcc cctactaggg
     2281 ggtgacagct ctaagaggcg ttaggcgatt aggctgaaaa tgtaggtcac accccagccg
     2341 tctgggaatg agcgagagag gagaagaggg actgagacac cgcacgggga aaaggtgaga
     2401 ggaagctgag ggctcatgag ggcattcaca gaagtgtggt ggtgattgga tttcagcttc
     2461 ttttagtctc cagatgaaag accagagttg ggatctccgc tttttctatc tcttctctcc
     2521 agctgggctg cttgtcttcg ggatgggcta ttaagagcga gtttcttcat ggaagctcat
     2581 ggcttatctt tctaatacag tgactctgtg aggaataaga cgagtcaggg tgaaggtagg
     2641 gcaggggctg aaatgtgagg atgcttgaaa agaagcagca gccccaacaa gctggctgag
     2701 gatgggctgg gaacgcgtat atgtctcctt gacacggagt cttaaaagag gttttgggta
     2761 gtatgtctgg actcttacca ctctaagctt tgagtgattg ggctgagaat tgtgtttcca
     2821 tggaggggtg gtgtttgggg tcaacatggc tctctatatt ggaaaccaag ctcacaacat
     2881 aagcaaggtc tgatatagga agcttctctg gggaagatgc cgtttgtttt tttccttttt
     2941 ggagaactga tagcacctgt ctttcactgg cttctgagtc cctgccccag gccaatgaga
     3001 tatgcaaata agcccttcat cttattggat gaggctcgag gaccacagat gatctcaggg
     3061 aaccagtcct ttaagagtgt cctctgggct ctgtgcccac agcccccttg ttctgaatac
     3121 agggacgaca ccagcccctg ctctatggag tatttagtct ccagggaagc tgtgcctttc
     3181 tggttctgac agtgactacg tcatctctgc cattacgaat tctgcagtcg acggtaccgc
     3241 gggcccggga tccaccggtc gccaccatgg cctcctccga gaacgtcatc accgagttca
     3301 tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag atcgagggcg
     3361 agggcgaggg ccgcccctac gagggccaca acaccgtgaa gctgaaggtg accaagggcg
     3421 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt
     3481 acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca
     3541 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggc gaccgtgacc caggactcct
     3601 ccctgcagga cggctgcttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg
     3661 acggccccgt gatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc
     3721 cccgcgacgg cgtgctgaag ggcgagaccc acaaggccct gaagctgaag gacggcggcc
     3781 actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct
     3841 actactacgt ggacgccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
     3901 agcagtacga gcgcaccgag ggccgccacc acctgttcct gtagcggccg cactcctcag
     3961 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     4021 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     4081 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     4141 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     4201 ttggtttaga gtttggcaac atatgccata tgctggctgc catgaacaaa ggtggctata
     4261 aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc
     4321 cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc
     4381 cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc
     4441 agtcatagct gtccctcttc tcttatgaag atccctcgac ctgcagccca agcttggcgt
     4501 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4561 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4621 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc
     4681 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4741 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4801 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4861 cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc
     4921 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4981 tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac
     5041 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5101 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5161 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5221 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5281 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5341 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5401 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     5461 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5521 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5581 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5641 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5701 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5761 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5821 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5881 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5941 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6001 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6061 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6121 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6181 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6241 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6301 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6361 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6421 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6481 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6541 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6601 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6661 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6721 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6781 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6841 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6901 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6961 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctg
//
