LOCUS       sequence-6915-ex        7807 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 7807)
  AUTHORS   Matsuda T, Cepko CL
  TITLE     Electroporation and RNA interference in the rodent retina in vivo 
            and in vitro.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22.
  PUBMED    14603031
REFERENCE   2  (bases 1 to 7807)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2004 Jan 6. 101(1):16-22."
COMMENT     Sequence Label: pCralbp-DsRed
FEATURES             Location/Qualifiers
     source          1..7807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             931..939
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      4063..4072
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4069..4746
                     /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(4249..4269)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     4661..4684
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4831..4850)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4896..4951
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4950..4969)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5310..5326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5310..5326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5323..5345)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5334..5350
                     /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(5358..5364,5365..5382,5383..5388))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5403..5424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5483..5678
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5529..5664
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5684..5818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5721..5740)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5775..5794
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5886..5903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6057..6645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6137..6156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6816..7607,7608..7676))
                     /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     7439..7458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7677..7781)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agtgatccct ttgctagcac tgagaccctt taatacgcct
       61 cctcatgttg tggggacccc ttaaccataa aattattttt gttgctccat cataactata
      121 attttgtcac ctttatgaat cataatgtaa atatctgata ttcaggatat ctgatatgca
      181 actcttgtga aagggtccat caaccacccc atcccccacc ctcagtgatc atgacccaca
      241 ggttgggtct aggcagccct ctcctgtaag actctggcca gccttagctt acaggggatt
      301 ccctgagtga acagcacaga gccgaaaatc gatgcaaaaa gcaagaggaa tttattgctc
      361 caacatgttg gggtcgccct gcacatgaag agagagaccg accacaggca gcccttcctt
      421 tacagcttct ccaggcagca gccattaggc acaatatgat tggcagaaca gtatgacttt
      481 taaactgatt ggtctatagg gaatgaggta ggtggccaac agtatctctg agcctcccct
      541 acccgcccgc aggtggccag tggttggtcc ctcctcccgg tctgaggaat gtaattagcc
      601 ccttccttcc ggaggggatg ggtgcgttct ttaagacctt tctcttccag gaggggaggg
      661 gtctggtgga attttctaaa gtacctgagc tgacctcttc actccaacta gaccatgtca
      721 tggtagtcct cttggtacac agggatgctt ggtgatgaat cacagagcct cttagcagct
      781 gggactctta cctcccatga gcatcacagc cagggaggtt ctttgggcat aacagcatgc
      841 cacagcctag gtgaattaac caatagaaaa tcattctctc ggaatctcgg tagttggagg
      901 tctaagatca aggtattatc ctttggtgtc acctccttat cttatagata aaccctgaac
      961 tgtggctgga tttcagagaa cgacctgtgc acatgagctc aaccctaccc tgagcacatg
     1021 gaggagagag aaccaggacc aaaggtgggc tgcttgggtc cagactgaag atgaatttaa
     1081 ttacagacaa ctatctgtgg tggaagtagg gatctcccgg acattggagg cccccacaga
     1141 gatctaaagg atgtgtatcc ctcatacccc ggtgataaac caaggcagag tctgggtgct
     1201 cctttgactg ttggatgcat tcccttagcc tgggtgctca ctggctctcc attcactggg
     1261 tacaactcag aactccccag aatacttccc agggattggg cagctaagcc cctgaagtgg
     1321 gtactttgca tctcaggact ttccccacag cttcagctgg ctctctagtc atcccagttc
     1381 tcagagggga ctgagtcatg tcgtggtgac ataaccacac tgaaaaaagt gcagtagcaa
     1441 ggacagaaag tcggacaagg gagatctgct gaggccccac acccatcccc tgcttccttg
     1501 cctttgcgag gtagagatcg gcagagatcg gagggtcagg gtacctagca agtcctccgc
     1561 tgctcagcct cgcccgctcc atttccctct gccacatggg acagtggacc ataggcctgg
     1621 gttgcagcaa tgatcacatt tataatttct cttcacttct ctatatgtcc tgccctttgt
     1681 gaagaaggag tagtctccca ggcccccctg cctcccagtt tgagaagcga gatggcaggg
     1741 aaatgagacg gggctggccc agatccagtg aagtggcttc cacatggctg aacagagagc
     1801 tccaagcagg agctcacaaa gcagggcttt gatggtagcc ccgcaggccg ggcctctccc
     1861 ctctccctca gggtgcttca cttgcttcat tctgggcctc ctgtgagcct gactgaatgg
     1921 gaaacggtgg gcctgagaag ttccttgtga cacactaatc ccagcttgct gacctgacca
     1981 cacccccaaa gaaagcagcc tcccgagctc cagaacaatt caggcatcca gtagccccca
     2041 agaaagagct gtcaggtaag tcagtaacaa ggcctggctg ggccagtgag tataagaaag
     2101 agtgtgtgca tgggagcggg gtaaatgatc agggatgggt ggcgaacacc acgtctcaga
     2161 ttgcttggac cggcctgtaa aggcgtacgc catttccgca gggtgagaca gaccgatcca
     2221 cgggcagacg caacacatac gaatgtggat tacctaactc aattagaatg gcaggtgatc
     2281 aaatactctt tatgccctag tattggtaat taattgtact ttatttgcta attctaaatg
     2341 ccggagaaca tcgatgtgga gataggggta gcatgttttt attacataca tgcctacagc
     2401 taatgggtaa gaacattttt aaaataagag ataagtgctt ctccgtgctg acgttaatct
     2461 tgcacttcta agatctggga cttgggctgc tgatctgcac atttggctag ggataatgca
     2521 ctaatacctc tcgagtcttg ctttgctttg tttgcttgag gcagggtttc tctacgtagc
     2581 cctagctatc ctggaactca ctatgaagaa cagactggct tcaaaatcag agattcccct
     2641 agacactggg attaaagatg cgtaccacta tacccagctt ttgcttttcg tttttttgag
     2701 atagagtctc atgtagcaca ggctggtttt gaattaacta tgtagccaag gataacctta
     2761 actaaggatt ctcttgcctc tatctttttt tttctttcct ttttaaaaat tactagatat
     2821 tttctttgtt tacatttcaa atgttatcgc ctttcctagt ttcctctcca aaagtcccct
     2881 ataccctcac ccccgccctg ctccccaacc cacccatttc tgtttcctgg ccctggcatt
     2941 cccctgtact ggggcatata atcttcgcaa gactaagggc ctctcctccc attgatggcc
     3001 aactaggcca tcttctgcta cataagcagc tagagacaca agctctgggg ggtactggtt
     3061 agttcatatt gttgttcctc ctatagggct gcagacccct tcagctcctt gggtactttc
     3121 tctagctcct tcgttgggga ccctgtgttc catcctatag atgactgtga gcgtccactt
     3181 ctgtatttgc caggcactgc ttttgcctct atcttgcaag ttctgggaat acagatgtgt
     3241 gccacaacag tcaactcttg gtctttacac ccacatatct ggatgtctgt ctactatcca
     3301 aagagatggc ttgtacttga ttcttgtggg tagatcccag cccccaagaa ttccctggtc
     3361 ctttgggggc tgtggtaggg aagggatagg ccctcagata ttctctgaga tcctcaagtt
     3421 tcagtgtaca aattgggtta atctcagtgc tagcaaaggg agaaaagact gttgtcgatc
     3481 aggagtcctc tctcggttat gtcacagaac cctgtggtcc tcacagcggt cacttttggt
     3541 agatattatt atccttcgtt ggctctcaat ctcagaagga ttaaatacct tgcctatggt
     3601 cacacaggtg gtaaaagtca gattcaaatt cagatctctc taacctataa ggtttctgtc
     3661 accagctccc ccagggactt tgcaaagaga tgacttggct tgaggcaaag agacttagtg
     3721 tcaggtggat acaggttcaa gtcctagctt aaataccaac tatgcgtaag tgaccgagct
     3781 gaccaatgga acctagccac aggcagaaac agaatgtaaa gatgttcaca gtcactgggg
     3841 acatttaggt cttggaggag aaaatctgga aatgtgggtc atgatggata tctgttctgg
     3901 tggcccagaa atctatgctg gtgtatatta tctaattaca tattaatcct tctatcccaa
     3961 caagggacgg tcaagactcg gatgaaatca cacaacttgc aagtgtgaga gacagcattg
     4021 caatccagga tcgaattccc gcgggcccgg gatccaccgg tcgccaccat ggcctcctcc
     4081 gagaacgtca tcaccgagtt catgcgcttc aaggtgcgca tggagggcac cgtgaacggc
     4141 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcca caacaccgtg
     4201 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     4261 ttccagtacg gctccaaggt gtacgtgaag caccccgccg acatccccga ctacaagaag
     4321 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     4381 gcgaccgtga cccaggactc ctccctgcag gacggctgct tcatctacaa ggtgaagttc
     4441 atcggcgtga acttcccctc cgacggcccc gtgatgcaga agaagaccat gggctgggag
     4501 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagac ccacaaggcc
     4561 ctgaagctga aggacggcgg ccactacctg gtggagttca agtccatcta catggccaag
     4621 aagcccgtgc agctgcccgg ctactactac gtggacgcca agctggacat cacctcccac
     4681 aacgaggact acaccatcgt ggagcagtac gagcgcaccg agggccgcca ccacctgttc
     4741 ctgtagcggc cgcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     4801 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     4861 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     4921 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     4981 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgcca tatgctggct
     5041 gccatgaaca aaggtggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     5101 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     5161 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     5221 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatga agatccctcg
     5281 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5341 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5401 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5461 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     5521 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5581 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5641 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     5701 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5761 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     5821 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5881 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5941 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6001 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6061 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6241 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6481 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6901 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6961 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7021 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7081 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7141 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7201 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7261 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7321 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7381 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7441 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7501 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7561 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7621 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7681 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7741 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7801 ccacctg
//
