LOCUS       sequence-469998-        7305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  backbone for makeTCR assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7305)
  TITLE     makeTCR collection
REFERENCE   2  (bases 1 to 7305)
  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: makeTCR_Lentiviral_Backbone
FEATURES             Location/Qualifiers
     source          1..7305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(171..191)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(589..600)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             778..1011
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(1037..1054)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1043..1267
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1385..1402)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1556..2144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1636..1655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2315..3106,3107..3175))
                     /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     2938..2957
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3176..3280)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3355..3374)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3546..3925
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3927..4125
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4080..4100
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4143..4323
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4370..4495
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4988..5221
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5406..5450
                     /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    5748..5865
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6063..7245
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6294..7236
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7193..7213
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      join(7302..7305,1..6)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ccatggagac gtgctaaccg tatcgcattg gccataccga agatggccgc gaagagaact
       61 agcccgggac catttaaatt gagccacgtc tcataacaac aatttcgtgc cagccacact
      121 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
      181 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
      241 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
      301 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
      361 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
      421 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
      481 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
      541 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
      601 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
      661 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
      721 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
      781 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
      841 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
      901 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
      961 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt
     1021 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1081 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1141 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1201 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     1261 tctatggcat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1321 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1381 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1441 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1501 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1561 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1621 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1681 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1741 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1801 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1861 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1921 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1981 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2041 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2101 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2161 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2221 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2281 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2341 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2401 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2461 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2521 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2581 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     2641 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2701 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     2761 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     2821 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     2881 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     2941 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3001 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3061 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3121 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3181 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3241 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3301 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     3361 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     3421 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     3481 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     3541 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     3601 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     3661 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     3721 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     3781 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     3841 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     3901 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     3961 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     4021 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     4081 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta
     4141 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     4201 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     4261 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     4321 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     4381 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     4441 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     4501 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     4561 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     4621 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     4681 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     4741 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     4801 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     4861 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     4921 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     4981 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     5041 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     5101 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     5161 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     5221 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     5281 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     5341 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     5401 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     5461 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     5521 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     5581 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     5641 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
     5701 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
     5761 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     5821 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     5881 gacagcagag atccagtttg gcgccatgtt gagtaattca tacaaaagga ctcgcccctg
     5941 ccttggggaa tcccagggac cgtcgttaaa ctcccactaa cgtagaaccc agagatcgct
     6001 gcgttcccgc cccctcaccc gcccgctctc gtcatcactg aggtggagaa gagcatgcgt
     6061 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     6121 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     6181 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     6241 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     6301 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     6361 ttacttccac gcccctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt
     6421 gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag
     6481 gcctggcttg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
     6541 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
     6601 ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt
     6661 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
     6721 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
     6781 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
     6841 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa
     6901 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
     6961 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     7021 ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     7081 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     7141 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     7201 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgacgttt agtgaaccgt
     7261 cagaaaattt aatacgactc actataagga gagtaattct agcca
//
