LOCUS       sequence-172972-        5106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mouse CAR cytoplasmic domain, SIV isoform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5106)
  TITLE     Rathjen lab plasmids
REFERENCE   2  (bases 1 to 5106)
  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-SIV
FEATURES             Location/Qualifiers
     source          1..5106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      30..485
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(117..136)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     327..348
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     611..633
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     625..642
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     626..642
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        803..1182
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1183..1386
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1337..1357
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1417..1461
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             1462..1485
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1932..2554
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2131..2150)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2578..2598)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2583..2912
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2763..2898
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2950..2969)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2951..2969)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2983..2999)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2983..2999)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2996..3018)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3007..3023
                     /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(3031..3037,3038..3055,3056..3061))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3076..3097
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3214..3231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3385..3973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3465..3484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4144..4935,4936..5004))
                     /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     4767..4786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5005..5106,1..3))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
       61 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
      121 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
      181 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
      241 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
      301 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
      361 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
      421 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
      481 aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      541 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      601 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct gatctataca
      661 ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt
      721 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
      781 tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
      841 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      901 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      961 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     1021 actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc
     1081 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct
     1141 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     1201 tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     1261 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaat
     1321 aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     1381 agagctcgtt tagtgaaccg tcagaattaa ttcaccatgt ctgcacttct gatcctagct
     1441 cttgttggag ctgcagttgc tgactacaaa gacgatgacg acaagcttgc cggaacgatc
     1501 gcgggcgccg tcatagggac gctgctggcc cttgtgctca tcggggccat cctcttctgc
     1561 tgtcacagga aacgcagaga agagaagtac gagaaggaag ttcatcatga tatcagggaa
     1621 gatgtgcctc ctccaaagag tcggacatcc actgccagga gctatattgg cagcaaccat
     1681 tcctccctgg gatccatgtc cccctctaac atggaggggt attccaagac gcagtataac
     1741 caagtcccca gtgaagactt tgaacgtgcg cctcagagcc cgactctggc acccgctaag
     1801 gtagctgccc ctaatctcag tcgaatggga gcggttcctg tgatgattcc tgcacagagc
     1861 aaggacgggt ctatagtata ggaattcatc gatagatctg atatcggtac cagtcgactc
     1921 tagaggatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     1981 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     2041 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     2101 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     2161 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     2221 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     2281 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     2341 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     2401 ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc
     2461 tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat
     2521 gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagct tggctgtgga
     2581 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2641 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2701 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2761 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2821 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2881 gaggcttttt tggaggccta ggcttttgca aaaagctcct cgaggaactg aaaaaccaga
     2941 aagttaattc cctatagtga gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt
     3001 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3061 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3121 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3181 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3241 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3301 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3361 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3421 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3481 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3541 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3601 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3661 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3721 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3781 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3841 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3901 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3961 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4021 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4081 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4141 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4201 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4261 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4321 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4381 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4441 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4501 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4561 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4621 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4681 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4741 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4801 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4861 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4921 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4981 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5041 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5101 ggttcc
//
