LOCUS       sequence-378823-        6975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  coexpression of Cre and tdT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6975)
  TITLE     Oka Lab Plasmids 2022
REFERENCE   2  (bases 1 to 6975)
  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"
COMMENT     Sequence Label: AV0.1-CWS-Cre-tdT
FEATURES             Location/Qualifiers
     source          1..6975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        341..644
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        645..848
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     799..819
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             916..936
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1978..2034
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2035..3465
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    3495..4083
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3548..3568)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3966..3977)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4104..4185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4141..4160)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4195..4214
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4238..4378
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4238..4367
                     /label=AAV2 ITR
     rep_origin      4453..4908
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4540..4559)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4750..4771
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4925..4944)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5044..5066
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5104..5122)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5190..5294
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5295..5363,5364..6155)
                     /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(5513..5532)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6326..6914
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6815..6834
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgtgg agtcgtgacg tgaattacgt catagggtta gggaggtcct
      181 gtattagagg tcacgtgagt gttttgcgac attttgcgac accatgtggt cacgctgggt
      241 atttaagccc gagtgagcac gcagggtctc catacgcgcc tagttattaa tagtaatcaa
      301 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      361 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      421 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      481 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      541 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      601 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      661 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      721 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      781 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      841 gcagagctgg tttagtgaac cgtcagatcc gctagcaagc ttggatcctt cgaaaccggt
      901 gctagcacca tggtgcccaa gaagaagagg aaagtctcca acctgctgac tgtgcaccaa
      961 aacctgcctg ccctccctgt ggatgccacc tctgatgaag tcaggaagaa cctgatggac
     1021 atgttcaggg acaggcaggc cttctctgaa cacacctgga agatgctcct gtctgtgtgc
     1081 agatcctggg ctgcctggtg caagctgaac aacaggaaat ggttccctgc tgaacctgag
     1141 gatgtgaggg actacctcct gtacctgcaa gccagaggcc tggctgtgaa gaccatccaa
     1201 cagcacctgg gccagctcaa catgctgcac aggagatctg gcctgcctcg cccttctgac
     1261 tccaatgctg tgtccctggt gatgaggaga atcagaaagg agaatgtgga tgctggggag
     1321 agagccaagc aggccctggc ctttgaacgc actgactttg accaagtcag atccctgatg
     1381 gagaactctg acagatgcca ggacatcagg aacctggcct tcctgggcat tgcctacaac
     1441 accctgctgc gcattgccga aattgccaga atcagagtga aggacatctc ccgcaccgat
     1501 ggtgggagaa tgctgatcca cattggcagg accaagaccc tggtgtccac agctggtgtg
     1561 gagaaggccc tgtccctggg ggttaccaag ctggtggaga gatggatctc tgtgtctggt
     1621 gtggctgatg accccaacaa ctacctgttc tgccgggtca gaaagaatgg tgtggctgcc
     1681 ccttctgcca cctcccaact gtccacccgg gccctggaag ggatctttga ggccacccac
     1741 cgcctgatct atggtgccaa ggatgactct gggcagagat acctggcctg gtctggccac
     1801 tctgccagag tgggtgctgc cagggacatg gccagggctg gtgtgtccat ccctgaaatc
     1861 atgcaggctg gtggctggac caatgtgaac attgtgatga actacatcag aaacctggac
     1921 tctgagactg gggccatggt gaggctgctc gaggatgggg acgttaacgg aagcggagcc
     1981 actaacttct ccctgttgaa acaagcaggg gatgtcgaag agaatcccgg gccaatggtg
     2041 agcaagggcg aggaggtcat caaagagttc atgcgcttca aggtgcgcat ggagggctcc
     2101 atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     2161 cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg
     2221 tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc accccgccga catccccgat
     2281 tacaagaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     2341 ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg acggcacgct gatctacaag
     2401 gtgaagatgc gcggcaccaa cttccccccc gacggccccg taatgcagaa gaagaccatg
     2461 ggctgggagg cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc
     2521 caccaggccc tgaagctgaa ggacggcggc cactacctgg tggagttcaa gaccatctac
     2581 atggccaaga agcccgtgca actgcccggc tactactacg tggacaccaa gctggacatc
     2641 acctcccaca acgaggacta caccatcgtg gaacagtacg agcgctccga gggccgccac
     2701 cacctgttcc tggggcatgg catcggcagc accggcagcg gcagctccgg caccgcctcc
     2761 tccgaggaca acaacatggc cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag
     2821 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     2881 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac
     2941 atcctgtccc cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc
     3001 cccgattaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     3061 gaggacggcg gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc
     3121 tacaaggtga agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag
     3181 accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc
     3241 gagatccacc aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc
     3301 atctacatgg ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg
     3361 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc
     3421 cgccaccacc tgttcctgta cggcatggac gagctgtaca agtaaggcgc gcccctgcag
     3481 ggaattctat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     3541 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     3601 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     3661 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     3721 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     3781 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     3841 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     3901 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     3961 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     4021 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     4081 cgcatcgata agtaagtgac tagaacttgt ttattgcagc ttataatggt tacaaataaa
     4141 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4201 tgtccaaact catctgcacg cgtggcgcgt cggccgcagg aacccctagt gatggagttg
     4261 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     4321 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     4381 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     4441 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4501 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     4561 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4621 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     4681 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4741 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     4801 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     4861 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     4921 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4981 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5041 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     5101 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5161 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5221 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5281 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5341 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5401 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5461 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5521 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5581 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5641 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5701 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5761 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5821 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5881 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5941 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6001 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6061 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6121 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6181 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6241 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6301 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6361 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6421 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6481 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6541 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6601 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6661 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6721 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6781 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6841 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6901 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6961 cttttgctca catgt
//
