LOCUS       sequence-172989-        5711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mouse CAR, TVV isoform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5711)
  TITLE     Rathjen lab plasmids
REFERENCE   2  (bases 1 to 5711)
  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: FTCAR2:pFlagCMV-mCAR-TVV
FEATURES             Location/Qualifiers
     source          1..5711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      26..481
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(113..132)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     323..344
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     607..629
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     621..638
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     622..638
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        799..1178
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1179..1382
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1333..1353
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1413..1457
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             1458..1481
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1485..1502
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     polyA_signal    2533..3155
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2732..2751)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3179..3199)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3184..3513
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3364..3499
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3551..3570)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3552..3570)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3584..3600)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3584..3600)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3597..3619)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3608..3624
                     /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(3632..3638,3639..3656,3657..3662))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3677..3698
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3815..3832)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3986..4574)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4066..4085)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4745..5536,5537..5605))
                     /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     5368..5387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5606..5710)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
       61 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      121 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      181 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      241 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
      301 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
      361 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
      421 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
      481 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
      541 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      601 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctgatc tatacattga
      661 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
      721 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
      781 caatatgacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      841 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      901 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      961 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     1021 cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg
     1081 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt
     1141 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     1201 ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     1261 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc
     1321 ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     1381 ctcgtttagt gaaccgtcag aattaattca ccatgtctgc acttctgatc ctagctcttg
     1441 ttggagctgc agttgctgac tacaaagacg atgacgacaa gctcctagta ccccgaggat
     1501 ctgcgaattc tttgagcatc actacacccg aacagaggat cgaaaaagcc aaaggggaaa
     1561 ctgcgtatct accatgcaag tttactctca gtcccgaaga ccagggacca ctggacattg
     1621 aatggctgat atccccgtct gataaccaga tagtggatca agtgatcatt ttgtattctg
     1681 gagacaaaat ttatgataac tactatccgg atctgaaagg acgggtacat tttacgagta
     1741 acgatgtcaa gtctggcgac gcatctataa atgtgaccaa cctgcagctg tcggacattg
     1801 gcacttacca gtgcaaagtg aagaaagccc ctggggttgc aaataagaaa ttcctgctga
     1861 ccgttcttgt taagccttca ggtacaagat gcttcgtgga tggatcggaa gagattggaa
     1921 atgacttcaa gctaaaatgt gaacccaagg aaggctccct tccactacag tttgaatggc
     1981 agaaactgtc ggactcccag acaatgccta cgccatggct ggcagaaatg acgtcaccag
     2041 ttatatctgt gaagaacgcc agttctgagt attctgggac atacagctgc acggttcaaa
     2101 acagagtggg ctctgaccag tgtatgctgc gactagacgt tgtcccaccc tccaaccgag
     2161 ccggaacgat cgcgggcgcc gtcataggga cgctgctggc ccttgtgctc atcggggcca
     2221 tcctcttctg ctgtcacagg aaacgcagag aagagaagta cgagaaggaa gttcatcatg
     2281 atatcaggga agatgtgcct cctccaaaga gtcggacatc cactgccagg agctatattg
     2341 gcagcaacca ttcctccctg ggatccatgt ccccctctaa catggagggg tattccaaga
     2401 cgcagtataa ccaagtcccc agtgaagact ttgaacgtgc gcctcagagc ccgactctgg
     2461 cacccgctaa gttcaagtac gcttacaaga ccgatggcat tacagtggta taagtcgact
     2521 ctagaggatc ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     2581 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     2641 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     2701 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     2761 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     2821 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     2881 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     2941 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     3001 cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg
     3061 ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca
     3121 tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc ttggctgtgg
     3181 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3241 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3301 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3361 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3421 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3481 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tcgaggaact gaaaaaccag
     3541 aaagttaatt ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg
     3601 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3661 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3721 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3781 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3841 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3901 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3961 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4021 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4081 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4141 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4201 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4261 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4321 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4381 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4441 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4501 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4561 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4621 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4681 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4741 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4801 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4861 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4921 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4981 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5041 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5101 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5161 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5221 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5281 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5341 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5401 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5461 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5521 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5581 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5641 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5701 gggttccgcg c
//
