LOCUS       sequence-172977-        5756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of chicken CAR, SIV isoform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5756)
  TITLE     Rathjen lab plasmids
REFERENCE   2  (bases 1 to 5756)
  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: FTCAR1:pFlagCMV-chCAR-SIV
FEATURES             Location/Qualifiers
     source          1..5756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..470
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(102..121)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     312..333
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     596..618
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     610..627
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     611..627
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        788..1167
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1168..1371
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1322..1342
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1402..1446
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             1447..1470
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1474..1491
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     polyA_signal    2567..3189
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2766..2785)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3213..3233)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3218..3547
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3398..3533
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3585..3604)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3586..3604)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3618..3634)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3618..3634)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3631..3653)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3642..3658
                     /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(3666..3672,3673..3690,3691..3696))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3711..3732
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3849..3866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4020..4608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4100..4119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4779..5570,5571..5639))
                     /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     5402..5421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5640..5744)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
       61 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
      121 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
      181 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
      241 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
      301 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
      361 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
      421 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca
      481 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
      541 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
      601 tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg
      661 caattagcca tattagtcat tggttatata gcataaatca atattggcta ttggccattg
      721 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
      781 ccatgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      841 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      901 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      961 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     1021 gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg
     1081 cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc
     1141 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt
     1201 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     1261 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg
     1321 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
     1381 aaccgtcaga attaattcac catgtctgca cttctgatcc tagctcttgt tggagctgca
     1441 gttgctgact acaaagacga tgacgacaag ctcctagtac cccgaggatc tgcgaattcc
     1501 ctaagtataa cttcagctga atcagcattt gaaaaagctc aaggagagag agttacgttg
     1561 ccatgtacct ttgaactctc agaagaggat gtaggcacac tagatattga gtgggttttg
     1621 ataccagcag atattcagaa gaaggaagaa acaataattc tatattctgg agataggatt
     1681 tataatcatt atcatcctgc tctggctggg cggctgcaat ttactagttc tgatcccaaa
     1741 tctggtgatg gttcagtgga tatcctgaat ttaaagtcag cagatactgg cacataccag
     1801 tgcaaagtga agaaggctcc tggagttgaa agcctaaaaa tacagttgaa cgtacttgta
     1861 aagccagcaa gcactaaatg ctccattgaa ggatcacagg agattggaaa ggacattgta
     1921 ttgaagtgtg catcacaaga aggaacccca cttttgtatt atgactggag aagagtagta
     1981 actggcacac aaggacttcc tgccacttcc gtactaaaca aaaatacagg ggaacttctc
     2041 ttgaaaaatg cctctaaaga ctattctggt acatacagtt gtgttgcttc aaaccgagtt
     2101 ggcacagatg aatgttctgt tgagctgaat gtcacccctc ctataaatac agctggtgta
     2161 attgctggag ctattctggg aactctgttg ggtcttgcct tgttggcttt tcttgtcatc
     2221 tgttgctgta agaagcatag agagaagaaa tatgagaaag aagtacatca tgaaatcaga
     2281 gaagatgttc tgcctccaaa aagtcgcagt tcaacagcac gcagctacat aggcagcaat
     2341 cgttcttctc tgggttcaat gtctccctca aatatggaag gatactccaa aactccatat
     2401 agccaggttc caagtgaaga ctttgaacgt acttctggtc aaaaccaaac cattgcatct
     2461 tcaaaggtag ctgcacctaa tttaagtaga atgggagctg tccctgtgat gattccagca
     2521 caaagcaaag atgggtccat agtatgagtc gactctagag gatcccgggt ggcatccctg
     2581 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     2641 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     2701 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     2761 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     2821 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     2881 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     2941 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3001 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     3061 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     3121 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     3181 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3301 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3361 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3421 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3481 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3541 ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat agtgagtcgt
     3601 attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3661 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3721 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3781 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3841 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3901 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3961 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4021 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4081 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4141 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4201 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4261 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4321 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4381 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4441 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4501 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4561 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4621 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4681 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4741 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4801 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4861 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4921 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4981 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5041 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5101 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5161 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5221 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5281 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5341 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5401 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5461 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5521 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5581 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5641 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5701 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccg
//
