LOCUS       sequence-386248-        8113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express single-chain peptide-MHC (CMV p5 epitope on HLA-A0101).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8113)
  AUTHORS   Yu B, Shi Q, Belk JA, Yost KE, Parker KR, Li R, Liu BB, Huang H, 
            Lingwood D, Greenleaf WJ, Davis MM, Satpathy AT, Chang HY
  TITLE     Engineered cell entry links receptor biology with single-cell 
            genomics.
  JOURNAL   Cell. 2022 Dec 22;185(26):4904-4920.e22. doi: 
            10.1016/j.cell.2022.11.016. Epub 2022 Dec 13.
  PUBMED    36516854
REFERENCE   2  (bases 1 to 8113)
  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"; doi: 
            "10.1016/j.cell.2022.11.016"; journalName: "Cell"; date: 
            "2022-12-22- 22"; volume: "185"; issue: "26"; pages: "4904-4920.e22"
COMMENT     Sequence Label: plentiCMV-sct-p5-b2m-HLA-A0101
FEATURES             Location/Qualifiers
     source          1..8113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1058..1284
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1285..1465
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1512..1637
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2130..2363
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2548..2592
                     /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    2859..2975
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2998..3301
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3302..3505
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3456..3476
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3502..3526
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3541..3550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    5023..5611
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5076..5096)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5494..5505)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5683..5916
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5994..6115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6031..6050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6085..6104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6155..6290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6310..6329)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6311..6329)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6339..6356)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6339..6355)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6348..6370)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6497..6952
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6584..6603)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6794..6815
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6978..7082
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7083..7151,7152..7943)
                     /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(7301..7320)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     1081 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     1141 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     1201 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     1261 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     1321 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1381 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1441 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     1501 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     1561 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1621 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1681 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1741 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1801 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1861 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1921 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1981 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     2041 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     2101 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     2161 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     2221 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     2281 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     2341 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     2401 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     2461 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     2521 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     2581 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     2641 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     2701 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     2761 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     2821 attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt
     2881 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2941 gaattacaaa aacaaattac aaaattcaaa attttatcga taagcttggg agttccgcgt
     3001 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     3061 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     3121 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     3181 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     3241 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     3301 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     3361 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     3421 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     3481 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagtgtaca
     3541 gccaccatgg cgacgggttc aagaacttcc ctacttcttg catttggcct gctttgtttg
     3601 ccgtggttac aggagggctc ggcaggatac tcagagcatc ctacattcac gagccaatat
     3661 ggatgcggag ggtccggagg gggaggtagc ggtggtggag gaagcatcca gcgtactcca
     3721 aagattcagg tttactcacg tcatccagca gagaatggaa agtcaaattt cctgaattgc
     3781 tatgtgtctg ggtttcatcc atccgacatt gaagttgact tactgaagaa tggagagaga
     3841 attgaaaaag tggagcattc agacttgtct ttcagcaagg actggtcttt ctatctcttg
     3901 tactacactg aattcacccc cactgaaaaa gatgagtatg cctgccgtgt gaaccatgtg
     3961 actttgtcac agcccaagat agttaagtgg gatcgagaca tgggaggtgg tggcagcggt
     4021 ggagggggct ccggcggtgg gggatccggc ggcggcggaa gcggctctca ctccatgagg
     4081 tatttcttca catccgtgtc ccggcccggc cgcggggagc cccgcttcat cgccgtgggc
     4141 tacgtggacg acacgcagtt cgtgcggttc gacagcgacg ccgcgagcca gaagatggag
     4201 ccgcgggcgc cgtggataga gcaggagggg ccggagtatt gggaccagga gacacggaat
     4261 atgaaggccc actcacagac tgaccgagcg aacctgggga ccctgcgcgg ctgctacaac
     4321 cagagcgagg acggttctca caccatccag ataatgtatg gctgcgacgt ggggccggac
     4381 gggcgcttcc tccgcgggta ccggcaggac gcctacgacg gcaaggatta catcgccctg
     4441 aacgaggacc tgcgctcttg gaccgcggcg gacatggcag ctcagatcac caagcgcaag
     4501 tgggaggcgg tccatgcggc ggagcagcgg agagtctacc tggagggccg gtgcgtggac
     4561 gggctccgca gatacctgga gaacgggaag gagacgctgc agcgcacgga cccccccaag
     4621 acacatatga cccaccaccc catctctgac catgaggcca ccctgaggtg ctgggccctg
     4681 ggcttctacc ctgcggagat cacactgacc tggcagcggg atggggagga ccagacccag
     4741 gacacggagc tcgtggagac caggcctgca ggggatggaa ccttccagaa gtgggcggct
     4801 gtggtggtgc cttctggaga ggagcagaga tacacctgcc atgtgcagca tgagggtctg
     4861 cccaagcccc tcaccctgag atgggagctg tcttcccagc ccaccatccc catcgtgggc
     4921 atcattgctg gcctggttct ccttggagct gtgatcactg gagctgtggt cgctgccgtg
     4981 atgtggagga ggaagagctc aggttgaacg cgttaagtcg acaatcaacc tctggattac
     5041 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     5101 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     5161 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     5221 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     5281 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     5341 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     5401 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     5461 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     5521 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     5581 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     5641 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5701 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     5761 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5821 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5881 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     5941 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     6001 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6061 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6121 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6181 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6241 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     6301 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     6361 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     6421 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     6481 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6541 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6601 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6661 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6721 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6781 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6841 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6901 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     6961 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     7021 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     7081 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     7141 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     7201 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     7261 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     7321 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     7381 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     7441 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     7501 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     7561 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7621 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7681 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7741 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     7801 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7861 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7921 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7981 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     8041 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8101 tcaaaggatc ttc
//
