LOCUS       sequence-172969-        5067 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mouse CAR cytoplasmic domain, TVV isoform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5067)
  TITLE     Rathjen lab plasmids
REFERENCE   2  (bases 1 to 5067)
  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: pFlagCMV-mCARcyt-TVV
FEATURES             Location/Qualifiers
     source          1..5067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        38..417
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        418..621
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     572..592
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             652..696
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             697..720
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1128..1750
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1327..1346)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1774..1794)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1779..2108
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1959..2094
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2146..2165)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2147..2165)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2179..2195)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2179..2195)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2192..2214)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2203..2219
                     /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(2227..2233,2234..2251,2252..2257))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2272..2293
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2410..2427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2581..3169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2661..2680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3340..4131,4132..4200))
                     /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     3963..3982
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4201..4305)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4332..4787
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4419..4438)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4629..4650
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4913..4935
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4927..4944
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4928..4944
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt gactagttat
       61 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      121 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      181 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      241 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg
      301 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      361 ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      421 atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca
      481 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      541 ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg
      601 gaggtctata taagcagagc tcgtttagtg aaccgtcaga attaattcac catgtctgca
      661 cttctgatcc tagctcttgt tggagctgca gttgctgact acaaagacga tgacgacaag
      721 cttgccggaa cgatcgcggg cgccgtcata gggacgctgc tggcccttgt gctcatcggg
      781 gccatcctct tctgctgtca caggaaacgc agagaagaga agtacgagaa ggaagttcat
      841 catgatatca gggaagatgt gcctcctcca aagagtcgga catccactgc caggagctat
      901 attggcagca accattcctc cctgggatcc atgtccccct ctaacatgga ggggtattcc
      961 aagacgcagt ataaccaagt ccccagtgaa gactttgaac gtgcgcctca gagcccgact
     1021 ctggcacccg ctaagttcaa gtacgcttac aagaccgatg gcattacagt ggtataagaa
     1081 ttcatcgata gatctgatat cggtaccagt cgactctaga ggatcccggg tggcatccct
     1141 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     1201 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     1261 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     1321 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     1381 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     1441 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     1501 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     1561 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac
     1621 tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg
     1681 gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc
     1741 ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa
     1801 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1861 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     1921 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     1981 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2041 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2101 tttgcaaaaa gctcctcgag gaactgaaaa accagaaagt taattcccta tagtgagtcg
     2161 tattaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2221 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2281 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2341 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2401 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2461 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2521 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2581 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2641 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2701 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2761 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2821 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2881 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2941 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3001 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3061 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3121 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3181 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3241 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3301 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3361 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3421 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3481 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3541 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3601 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3661 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3721 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3781 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3841 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3901 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3961 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4021 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4081 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4141 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4201 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4261 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4321 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4381 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4441 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4501 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4561 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4621 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4681 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4741 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc
     4801 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     4861 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     4921 cgacgttgta aaacgacggc cagtgccaag ctgatctata cattgaatca atattggcaa
     4981 ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     5041 acgttgtatc tatatcataa tatgtac
//
