LOCUS       sequence-311549-        6989 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 6989)
  AUTHORS   Marshel JH, Mori T, Nielsen KJ, Callaway EM
  TITLE     Targeting single neuronal networks for gene expression and cell 
            labeling in vivo.
  JOURNAL   Neuron. 2010 Aug 26. 67(4):562-74.
  PUBMED    20797534
REFERENCE   2  (bases 1 to 6989)
  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. 
            2010 Aug 26. 67(4):562-74."
COMMENT     Sequence Label: pCAG-YTB
FEATURES             Location/Qualifiers
     source          1..6989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(1066..1068,1069..1071,1072..1782)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1111..1132)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1372..1391)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1719..1740
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1792..1857
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             2338..2391
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     polyA_signal    4019..4153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4043..4062)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4097..4116
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4266..4286)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4266..4284)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4305..4321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4305..4321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4318..4340)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4329..4345
                     /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(4353..4359,4360..4377,4378..4383))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4398..4419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4536..4553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4707..5295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4787..4806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5466..6257,6258..6326))
                     /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     6089..6108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6327..6431)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6457..6912)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6594..6615)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6806..6825
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat ccaccatggt gagcaagggc
     1081 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1141 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1201 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1261 ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1321 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1381 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1441 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1501 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     1561 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     1621 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1681 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1741 accgccgccg ggatcactct cggcatggac gagctgtaca agggtagtgg tgtgaaacag
     1801 actttgaatt ttgaccttct caagttggcg ggagacgtgg agtccaaccc agggcccatg
     1861 gcgcggctgc tgcccgcgct gctgctgctg ctgctgcccg gtaacgtgac cggtaacggg
     1921 tccggtaacg gttctttgtc ccgttgcccc cccggtcagt tccgctgctc ggagccgccc
     1981 ggtgcccacg gggagtgtta cccgcaggac tggctgtgcg acggacaccc cgactgcgac
     2041 gacgggcggg acgagtgggg ctgcgggacc agcgcgatcc ccgcggtgcc cacggacaac
     2101 ggcacagagg ctcccactgc ccctgctcct ggacgtgctc tgccagccag gaatcacggc
     2161 cgcatgtgga tgctgatcac tgcagtgctc ctgtgctgcc tggtagctgt gggtggtatc
     2221 gctgcatggg ggaagtccaa agcaaaaagc aggtctgaca tcttcagtct tgaaagcgca
     2281 tccaaggagc tgctggtgcc tgacaagagc caggcagact tgttctccgg tagtggagag
     2341 ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga atcctggccc aatggttcct
     2401 caggctctcc tgtttgtacc ccttctggtt tttccattgt gttttgggaa attccctatt
     2461 tacacgatac cagacaagct tggtccctgg agtccgattg acatacatca cctcagctgc
     2521 ccaaacaatt tggtagtgga ggacgaagga tgcaccaacc tgtcagggtt ctcctacatg
     2581 gaacttaaag ttggatacat cttagccata aaagtgaacg ggttcacttg cacaggcgtt
     2641 gtgacggagg ctgaaaccta cactaacttc gttggttatg tcacaaccac gttcaaaaga
     2701 aagcatttcc gcccaacacc agatgcatgt agagccgcgt acaactggaa gatggccggt
     2761 gaccccagat atgaagagtc tctacacaat ccgtaccctg actaccgctg gcttcgaact
     2821 gtaaaaacca ccaaggagtc tctcgttatc atatctccaa gtgtggcaga tttggaccca
     2881 tatgacagat cccttcactc gagggtcttc cctagcggga agtgctcagg agtagcggtg
     2941 tcttctacct actgctccac taaccacgat tacaccattt ggatgcccga gaatccgaga
     3001 ctagggatgt cttgtgacat ttttaccaat agtagaggga agagagcatc caaagggagt
     3061 gagacttgcg gctttgtaga tgaaagaggc ctatataagt ctttaaaagg agcatgcaaa
     3121 ctcaagttat gtggagttct aggacttaga cttatggatg gaacatgggt ctcgatgcaa
     3181 acatcaaatg aaaccaaatg gtgccctccc gataagttgg tgaacctgca cgactttcgc
     3241 tcagacgaaa ttgagcacct tgttgtagag gagttggtca ggaagagaga ggagtgtctg
     3301 gatgcactag agtccatcat gacaaccaag tcagtgagtt tcagacgtct cagtcattta
     3361 agaaaacttg tccctgggtt tggaaaagca tataccatat tcaacaagac cttgatggaa
     3421 gccgatgctc actacaagtc agtcagaact tggaatgaga tcctcccttc aaaagggtgt
     3481 ttaagagttg gggggaggtg tcatcctcat gtgaacgggg tgtttttcaa tggtataata
     3541 ttaggacctg acggcaatgt cttaatccca gagatgcaat catccctcct ccagcaacat
     3601 atggagttgt tggaatcctc ggttatcccc cttgtgcacc ccctggcaga cccgtctacc
     3661 gttttcaagg acggtgacga ggctgaggat tttgttgaag ttcaccttcc cgatgtgcac
     3721 aatcaggtct caggagttga cttgggtctc ccgaactggg ggaagtatgt attactgagt
     3781 gcaggggccc tgactgcctt gatgttgata attttcctga tgacatgttg tagaagagtc
     3841 aatcgatcag aacctacgca acacaatctc agagggacag ggagggaggt gtcagtcact
     3901 ccccaaagcg ggaagatcat atcttcatgg gaatcacaca agagtggggg tgagaccaga
     3961 ctgtaaggcg cgccgtttcc tctcgagcct ctagaactat agtgagtcgt attacgtaga
     4021 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4081 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4141 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     4201 gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt
     4261 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     4321 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt
     4381 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4441 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4501 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4561 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4621 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4681 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4741 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4801 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4861 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4921 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4981 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5041 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5101 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5161 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5221 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5281 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5341 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5401 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5461 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5521 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5581 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5641 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5701 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5761 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5821 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5881 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5941 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6001 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6061 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6121 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6181 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6241 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6301 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6361 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6421 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     6481 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     6541 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     6601 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     6661 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     6721 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     6781 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     6841 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     6901 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     6961 gggcctcttc gctattacgc cagtcgacc
//
