LOCUS       sequence-6913-ex        6090 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 6090)
  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 6090)
  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: pRho-DsRed
FEATURES             Location/Qualifiers
     source          1..6090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2346..2355
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2352..3029
                     /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     2944..2967
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3114..3133)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3179..3234
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3233..3252)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3593..3609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3593..3609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3606..3628)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3617..3633
                     /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(3641..3647,3648..3665,3666..3671))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3686..3707
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3766..3961
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3812..3947
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3967..4101
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4004..4023)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4058..4077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4169..4186)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4340..4928)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4420..4439)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5099..5890,5891..5959))
                     /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     5722..5741
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5960..6064)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agccagcttc cacgttgtca gaagcgttag cgggtaacaa
       61 gcaaggccat gagtttgagg gtcgggggga cagcctggcc agggcagggc tgcgtgctct
      121 ggtcttgacc cagggagtcc atcgtctgtc cctctgtctg gccaccaggg gcgtgttaga
      181 atcaaactct tcaccttgac ctctttagcg atggacctaa tcacagggcc aagcaccttg
      241 agcagctgcg ccccacccac ccgccacacc tgcctgcccc ttgctccttc cccccagagc
      301 acttaccacc aggctcccca cacaagtcct gatgctgccc gctgcccggg acagcagggc
      361 tctggagccc aggtgggctg cagcgaaggg agcacgtgtc tgagcagaga ggtgtagaca
      421 tcccaggtgg taagagggac agcacaaggg agcctcagcg atggaggtct cagacactgg
      481 gatgcttcct ggccaccctt gcgttcccca cctcggggcc tttgcactgg ccgctctctc
      541 tccttggaca gctccacccc ttagtgctcc ttccatcctc gagttctcag agaggccttc
      601 ctgggcggct ccaggcaact gtgtgtcggg aggacgtcac catctgaaca gtccctgtgg
      661 tcgtttgttt catggtttgt gggggcccct ccccgccccc ccagaagaag gacacgtcct
      721 acaggcaggg gcttggtcct tcttgtccac tgtgtccccc agaactgagc gtcttgtgta
      781 gaacacagtc ggtgccagat aaaactcaca gctggactgt ggtcattgag gaccccgaag
      841 gccaggctgc ccgcgacccc acttcactcc taggaggggt cttcacgatc ctattgtaga
      901 tgggggccct tgttcgcacc ctgagctcgt gctgctgttc tgggatacgc catcacggct
      961 tgccgatcgc acggcacccg aggcccacgg ccctaaccca gcctgacccc agtcctggcc
     1021 tcggcccgtg tgctgttccc agggcccacc tcccctccat atgggagtgc ccgggaccac
     1081 acctccttgc tcaccgcagg gatctggtgc ccggccaagg cttggccccg ttcacctcct
     1141 ggagacgagg ccttgcttca ggagccctaa gagcttctgt cccctccccg ctttggttcc
     1201 ctctgccctg gtcccttcct ccaccatcgc tcccttcagc tcatccttga gggcttttga
     1261 gccaccttcc tctccccaag atctgcccaa gttcccctgg ataaagccct ggcactaagc
     1321 cccaccccag ctgaccctag cccacgaggg cccacatccc agatgggcct cgggccttca
     1381 ccgggcctct gtcttcagtc tatgctgact tccccacttg gcaacctggc aaagtgcccc
     1441 cctccctcaa cacattgtcc ccctggacgt ctctgcagat aggtggcctc cccgccccgg
     1501 gtccagagcg ggtcttccca cacggaagtc ctcctcagcc cataagctcc tccttcctgg
     1561 agcctcgggt gccccagtgg cctgcacata aatgtcggtt tgaacaagtg agtgagtgaa
     1621 ctccagttca agggaacaaa tgctttttga gctcctgggc aacgggggct tgtgtccggt
     1681 ccccggagcg ccccgctgca cttgtgaagc tcacgacctg gccggtgggc actgtcagtg
     1741 tgtggtgact ggataagccc agaggtgacc caggtgggga agggacagct caggggctga
     1801 gtgaggggat gaccgaagga ccagcgctct gccctccagg agctcgtggg ccaggcggga
     1861 ggaaggcaaa atttgcttgc cctgagccag agggaagtgg gcctagagct aggggtccca
     1921 atgccaggcc aggcccatca gctgagatgc cagaggggga caggggagtt gggggcaggc
     1981 ctctgctctt tcccagggtc cccagcacgc cccgccttct ccccgaccac agccttctac
     2041 ctggagtcac cttggcccca cctggaagcc aattaagccc ctcgttgcag cagtgaggat
     2101 taatatgatt aataacgccc ccaatctccg aggtgctgat tcagccggga gcttagggag
     2161 gggaggtcac ttcataaggg cttggggggg gagttggagc cacgagtcgt ccagccggag
     2221 ccccgtgtgg ctgagctccg gcctcagaag catccccggg ttcgcgccgc cggggatcct
     2281 ctagagtcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac
     2341 cggtcgccac catggcctcc tccgagaacg tcatcaccga gttcatgcgc ttcaaggtgc
     2401 gcatggaggg caccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2461 cctacgaggg ccacaacacc gtgaagctga aggtgaccaa gggcggcccc ctgcccttcg
     2521 cctgggacat cctgtccccc cagttccagt acggctccaa ggtgtacgtg aagcaccccg
     2581 ccgacatccc cgactacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     2641 tgaacttcga ggacggcggc gtggcgaccg tgacccagga ctcctccctg caggacggct
     2701 gcttcatcta caaggtgaag ttcatcggcg tgaacttccc ctccgacggc cccgtgatgc
     2761 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc
     2821 tgaagggcga gacccacaag gccctgaagc tgaaggacgg cggccactac ctggtggagt
     2881 tcaagtccat ctacatggcc aagaagcccg tgcagctgcc cggctactac tacgtggacg
     2941 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggagcag tacgagcgca
     3001 ccgagggccg ccaccacctg ttcctgtagc ggccgcactc ctcaggtgca ggctgcctat
     3061 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     3121 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     3181 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     3241 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     3301 gcaacatatg ccatatgctg gctgccatga acaaaggtgg ctataaagag gtcatcagta
     3361 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     3421 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     3481 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     3541 tcttctctta tgaagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc
     3601 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3661 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     3781 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3841 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3901 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3961 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4021 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4081 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4141 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4201 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4261 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4321 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4381 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4441 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4501 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     4561 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4621 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4681 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4741 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     4801 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4861 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4921 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4981 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5041 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5101 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5161 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5221 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5281 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5341 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5401 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5461 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5521 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5581 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5641 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5701 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5761 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5821 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5881 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5941 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6001 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6061 cgcgcacatt tccccgaaaa gtgccacctg
//
