LOCUS       sequence-358564-        5613 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 5613)
  AUTHORS   Wickersham IR, Lyon DC, Barnard RJ, Mori T, Finke S, Conzelmann KK, 
            Young JA, Callaway EM
  TITLE     Monosynaptic restriction of transsynaptic tracing from single, 
            genetically targeted neurons.
  JOURNAL   Neuron. 2007 Mar 1. 53(5):639-47.
  PUBMED    17329205
REFERENCE   2  (bases 1 to 5613)
  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: "Neuron. 
            2007 Mar 1. 53(5):639-47."
COMMENT     Sequence Label: pCAG-DsRed2
FEATURES             Location/Qualifiers
     source          1..5613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          117..1133
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1057..1079
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1141..1160
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    1226..1278
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1286..1295
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1292..1969
                     /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(1472..1492)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1884..1907
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        complement(2008..2026)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2009..2026)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2046..2063)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2052..2163
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2267..2286)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2268..2286)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2293..2310)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2293..2309)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2302..2324)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2451..2906
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2538..2557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2748..2769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2932..3036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3037..3105,3106..3897)
                     /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     complement(3255..3274)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4068..4656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4557..4576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4810..4827
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4944..4965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4980..4985,4986..5003,5004..5010)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5018..5034
                     /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)."
     primer_bind     5023..5045
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5042..5058
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5042..5058
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5079..5097
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        5121..5500
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
       61 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      121 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      181 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      241 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      301 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt
      361 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg
      421 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg
      481 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
      541 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc
      601 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg
      661 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc
      721 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct
      781 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
      841 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
      901 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
      961 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
     1021 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1081 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1141 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgtgg cggccgcctg
     1201 ggccggtgac cgtaccgggc cccccctcga gctcaagctt cgaattctgc agtcgacggt
     1261 accgcgggcc cgggatccac cggtcgccac catggcctcc tccgagaacg tcatcaccga
     1321 gttcatgcgc ttcaaggtgc gcatggaggg caccgtgaac ggccacgagt tcgagatcga
     1381 gggcgagggc gagggccgcc cctacgaggg ccacaacacc gtgaagctga aggtgaccaa
     1441 gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttccagt acggctccaa
     1501 ggtgtacgtg aagcaccccg ccgacatccc cgactacaag aagctgtcct tccccgaggg
     1561 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggcgaccg tgacccagga
     1621 ctcctccctg caggacggct gcttcatcta caaggtgaag ttcatcggcg tgaacttccc
     1681 ctccgacggc cccgtgatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct
     1741 gtacccccgc gacggcgtgc tgaagggcga gacccacaag gccctgaagc tgaaggacgg
     1801 cggccactac ctggtggagt tcaagtccat ctacatggcc aagaagcccg tgcagctgcc
     1861 cggctactac tacgtggacg ccaagctgga catcacctcc cacaacgagg actacaccat
     1921 cgtggagcag tacgagcgca ccgagggccg ccaccacctg ttcctgtagc ggccgccacc
     1981 gcggtggagc tgtttaaacg gccctattct atagtgtcac ctaaatgcta gagctcgctg
     2041 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2101 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2161 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2221 gggggaggat tgggaagaca atagcgcggc cgctctagcc aattcgccct atagtgagtc
     2281 gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2341 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2401 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     2461 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2521 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2581 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2641 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2701 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2761 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2821 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2881 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     2941 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3001 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3061 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3121 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3181 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3241 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3301 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3361 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3421 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3481 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3541 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3601 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3661 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3721 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3781 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3841 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3901 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3961 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4021 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4081 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4141 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4201 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4261 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4321 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4381 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4441 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4501 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4561 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4621 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4681 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4741 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4801 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     4861 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     4921 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     4981 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5041 acaggaaaca gctatgacca tgattacgcc aagctcgaaa ttaaccctca ctaaagggaa
     5101 caaaagctgg agctccactc gacattgatt attgactagt tattaatagt aatcaattac
     5161 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     5221 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     5281 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     5341 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     5401 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     5461 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
     5521 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
     5581 ttaattattt tgtgcagcga tgggggcggg ggg
//
