LOCUS       sequence-382464-        7542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre/LoxP-dependent expression of a tdTomato. Expression of Cre 
            excises a STOP codon before tdTomato, facilitating fluorescence 
            expression of tdTomato..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7542)
  TITLE     Bordey Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7542)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCALNL-tdTomato
FEATURES             Location/Qualifiers
     source          1..7542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(15..119)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        150..529
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          807..1823
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1747..1769
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1831..1850
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1898..1931)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1981..2775
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2035..2054)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2645..2664
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2955..3089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3103..3136)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3208..3217
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3214..4644
                     /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
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(4730..4749)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4795..4850
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4849..4868)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5211..5227)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5211..5227)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5224..5246)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5235..5251
                     /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(5259..5265,5266..5283,5284..5289))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5304..5325
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      5410..5544
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5564..5698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5766..5783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5937..6525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6017..6036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6696..7487,7488..7542,1..14))
                     /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     7319..7338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
       61 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      121 acatttcccc gaaaagtgcc acctgggtcg acattgatta ttgactagtt attaatagta
      181 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      241 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      301 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      361 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      421 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      481 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
      541 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
      601 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgccrg
      661 gggggggggg gggggggggg gggsggggsg gggsgrggcg gagaggtgcg gcggcagcca
      721 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct
      781 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc
      841 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
      901 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc
      961 ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg
     1021 gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct
     1081 ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca
     1141 gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg
     1201 ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt
     1261 cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt
     1321 cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc
     1381 ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggrgggc tcgggggagg
     1441 ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc
     1501 ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc
     1561 cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg
     1621 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     1681 tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca
     1741 gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc
     1801 atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc
     1861 attttggcaa agaattgatt aattcgagcg aacgcgtata acttcgtata gcatacatta
     1921 tacgaagtta tctcgagtcg gatttgatct gatcaagaga caggatgagg atcgtttcgc
     1981 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     2041 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     2101 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     2161 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     2221 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     2281 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     2341 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     2401 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     2461 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     2521 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     2581 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     2641 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     2701 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     2761 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     2821 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     2881 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     2941 cccaccccat cgataacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3001 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3061 tcatcaatgt atcttatcat gtctggatca atccgaacgc gtataacttc gtatagcata
     3121 cattatacga agttatctcg agtcgctcgg tacgatttaa attgaattct gcagtcgacg
     3181 gtaccgcggg cccgggatcc accggtcgcc accatggtga gcaagggcga ggaggtcatc
     3241 aaagagttca tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca cgagttcgag
     3301 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     3361 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt catgtacggc
     3421 tccaaggcgt acgtgaagca ccccgccgac atccccgatt acaagaagct gtccttcccc
     3481 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc
     3541 caggactcct ccctgcagga cggcacgctg atctacaagg tgaagatgcg cggcaccaac
     3601 ttcccccccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag
     3661 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc accaggccct gaagctgaag
     3721 gacggcggcc actacctggt ggagttcaag accatctaca tggccaagaa gcccgtgcaa
     3781 ctgcccggct actactacgt ggacaccaag ctggacatca cctcccacaa cgaggactac
     3841 accatcgtgg aacagtacga gcgctccgag ggccgccacc acctgttcct ggggcatggc
     3901 accggcagca ccggcagcgg cagctccggc accgcctcct ccgaggacaa caacatggcc
     3961 gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag
     4021 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     4081 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg
     4141 tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc
     4201 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc
     4261 gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc
     4321 accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     4381 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag
     4441 ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc
     4501 gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag
     4561 gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgtac
     4621 ggcatggacg agctgtacaa gtaggcggcc gcactcctca ggtgcaggct gcctatcaga
     4681 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     4741 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     4801 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     4861 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     4921 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     4981 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     5041 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     5101 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     5161 ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg
     5221 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5281 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5341 ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag
     5401 caaccatagt cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5461 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5521 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     5581 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5641 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     5701 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5761 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5821 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5881 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5941 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6001 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6061 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6121 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6181 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6241 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6301 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6361 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6421 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6481 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6541 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6601 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6661 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6721 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6781 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6841 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6901 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6961 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7021 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7081 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7141 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7201 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7261 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7321 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7381 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7441 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7501 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aa
//
