LOCUS       sequence-390333-        7190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LV Expression of CD3D cDNA with mutation at c.209T>C to assess a 
            bystander edit .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7190)
  TITLE     Kohn lab plasmids
REFERENCE   2  (bases 1 to 7190)
  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: pCCL-c-MNDU3-CD3D A0 cDNA
FEATURES             Location/Qualifiers
     source          1..7190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..39
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        21..39
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        140..519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        520..723
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     674..694
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             738..918
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    965..1090
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1583..1816
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2001..2045
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2338..2455
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      3143..3152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             3775..4008
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4080..4201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4117..4136)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4171..4190
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4241..4376
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4396..4415)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4397..4415)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4425..4442)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4425..4441)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4434..4456)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4583..5038
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4670..4689)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4880..4901
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5064..5168
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5169..5237,5238..6029)
                     /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     complement(5387..5406)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6200..6788
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6689..6708
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6942..6959
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7076..7097
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7112..7117,7118..7135,7136..7142)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7150..7166
                     /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)."
     primer_bind     7155..7177
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7174..7190
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7174..7190
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa
       61 gcttggccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt
      121 ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg
      181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      361 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      481 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      541 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      601 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      661 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      721 gctcgtttag tgaaccgggg tctctctggt tagaccagat ctgagcctgg gagctctctg
      781 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
      841 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
      901 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
      961 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     1021 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     1081 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     1141 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     1201 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     1261 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     1321 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     1381 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     1441 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     1501 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     1561 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     1621 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     1681 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     1741 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     1801 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     1861 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     1921 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     1981 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     2041 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     2101 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     2161 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg
     2221 acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat
     2281 tagtgaacgg atctcgacgg tatcgatctc gacacaaatg gcagtattca tccacaattt
     2341 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     2401 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     2461 ttattacagg gacagcagag atccagtttg ggtcgaggat atcgagatcg attagtccaa
     2521 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag
     2581 ctgaagccta tagagtacga gccatagata gaataaaaga ttttatttag tctccagaaa
     2641 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc taggatcaag gttaggaaca
     2701 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     2761 cagggccaag aacagttgga acagcagaat atgggccaaa caggatatct gtggtaagca
     2821 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
     2881 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     2941 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     3001 gagctcaata aaagagccca caacccctca ctcggcgcga tcgatgaatt cgagctcggt
     3061 acccggggat cccgggtgat cagtcgagct caagcttcga attctgcagt cgacggtacc
     3121 gcgggcccgg gatccaccgg tcgccaccat ggagcacagc accttcctgt ctggcctggt
     3181 actggctacc cttctctcgc aagtgagccc cttcaagata cctatagagg aacttgagga
     3241 cagagtgttt gtgaattgca ataccagcat cacatgggta gagggaacgg tgggaacact
     3301 gctctcagac attacaagac tggacctggg aaaacgcatc ctggacccac gaggaacata
     3361 taggtgtaat gggacagata tatacaagga caaagaatct accgtgcaag ttcattatcg
     3421 aatgtgccag agctgtgtgg agctggatcc agccaccgtg gctggcatca ttgtcactga
     3481 tgtcattgcc actctgctcc ttgctttggg agtcttctgc tttgctggac atgagactgg
     3541 aaggctgtct ggggctgccg acacacaagc tctgttgagg aatgaccagg tctatcagcc
     3601 cctccgagat cgagatgatg ctcagtacag ccacctgggc ggcaactggg ctcggaacaa
     3661 gtgataaagc ggccaactcg acgggcccgc ggaattcgag ctcggtacct ttaagaccaa
     3721 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     3781 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     3841 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     3901 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     3961 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     4021 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     4081 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4141 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4201 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
     4261 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4321 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4381 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     4441 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     4501 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4561 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4621 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4681 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4741 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4801 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4861 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4921 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4981 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     5041 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5101 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5161 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5221 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5281 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5341 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5401 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5461 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5521 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5581 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5641 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5701 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5761 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5821 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5881 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5941 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6001 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6061 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6121 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6181 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6241 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6301 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6361 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6421 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6481 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6541 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6601 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6661 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6721 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6781 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6841 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6901 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6961 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     7021 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     7081 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     7141 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
//
