LOCUS       sequence-446461-        8669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid expressing human TUBA1B protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8669)
  TITLE     Lentiviral plasmids expressing popular genes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8669)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pLenti puro HA-TUBA1B
FEATURES             Location/Qualifiers
     source          1..8669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     49..69
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        51..69
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        97..323
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             324..504
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    551..676
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1169..1402
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1587..1631
                     /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"
     primer_bind     1929..1949
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        2238..2441
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2392..2412
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2489..2515
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3956..4073
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4122..4632
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4551..4569
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             4654..5253
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4654..4673)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5150..5170
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5381..5614
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5686..5807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5723..5742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5777..5796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5847..5982
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6002..6021)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6003..6021)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6053..6070)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6053..6069)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6062..6084)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6278..6297)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6397..6419
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6457..6475)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6543..6647
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6648..6716,6717..7508)
                     /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(6866..6885)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7679..8267
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8168..8187
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8421..8438
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8555..8576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8591..8596,8597..8614,8615..8621)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8629..8645
                     /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     8634..8656
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8653..8669
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8653..8669
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgattacg ccaagcttgc atgcctgcag catgattacg ccaagcgcgc aattaaccct
       61 cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt
      121 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc
      181 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga
      241 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta
      301 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc
      361 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
      421 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
      481 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
      541 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
      601 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
      661 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
      721 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
      781 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
      841 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
      901 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
      961 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
     1021 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     1081 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     1141 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     1201 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     1261 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     1321 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     1381 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     1441 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     1501 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     1561 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     1621 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     1681 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     1741 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     1801 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     1861 ttcgattagt gaacggatct cgacggtatc gatcacgaga ctagcctcga gcggccgccc
     1921 ccttcaccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     1981 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     2041 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     2101 actatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2161 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2221 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2281 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2341 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2401 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     2461 tccgctagcg ctaccggtcg ccaccatgta cccatacgat gttccggatt acgctagcgg
     2521 atccatgcgg gagtgcatca gtatccacgt aggtcaggca ggcgtacaaa taggcaatgc
     2581 ttgctgggag ctttactgcc ttgagcatgg aatccaaccg gatggacaaa tgccatctga
     2641 taagaccatc ggaggaggcg acgattcttt taacacattc ttttccgaga cgggggctgg
     2701 taagcatgtc ccgcgagcgg tgtttgttga cttggagcca acagtgatcg atgaagttcg
     2761 aaccggcact tacagacaac ttttccatcc cgaacaactg ataaccggaa aagaggatgc
     2821 tgcgaataac tacgcgagag gtcattatac catcggcaaa gaaataatcg acctggttct
     2881 ggatcgaatc cggaagctgg ccgatcagtg tacagggctc cagggtttcc tggtgttcca
     2941 tagtttcggg ggaggtactg ggtcaggttt tacgtctttg ctcatggagc ggctcagcgt
     3001 tgattatggg aagaaatcta aattggaatt cagcatctat ccggccccgc aggtatctac
     3061 tgcagtcgta gagccatata acagtatact tacgactcac actacgcttg aacattctga
     3121 ctgtgccttc atggtagata atgaagccat ttatgatata tgcaggcgca atctcgacat
     3181 cgaacgaccg acttacacaa acctcaatag gctgatatcc cagatagtct cctctatcac
     3241 cgcctccctt agattcgacg gtgcactcaa cgtagacctc acagaattcc agacaaattt
     3301 ggtcccctac ccgaggatcc actttccact tgctacatac gcccccgtca tctctgcaga
     3361 gaaagcatac cacgagcaac tcagcgtcgc ggaaataact aatgcgtgtt tcgaaccggc
     3421 gaaccagatg gtcaagtgtg accccaggca tggtaaatat atggcctgtt gcttgttgta
     3481 ccgcggcgac gttgtaccaa aagacgtcaa cgccgccata gcaaccatta agactaagcg
     3541 ctcaatccaa tttgtagact ggtgtccaac ggggtttaag gtcggaataa attaccaacc
     3601 tccgacggta gtgccaggtg gcgacttggc aaaggttcag cgcgcagtct gcatgcttag
     3661 taacacgacg gctattgctg aagcttgggc ccgccttgac cataaatttg atttgatgta
     3721 tgccaaaagg gcgtttgtgc actggtatgt aggggagggg atggaagagg gtgagtttag
     3781 cgaggcacga gaagacatgg cagctcttga aaaagattac gaagaagttg gggtggattc
     3841 tgtggagggt gagggggagg aggagggtga agagtactag ccggactcag atctcgagct
     3901 caagcttctc tagagaattc tcgacctcga gacaaatggc agtattcatc cacaatttta
     3961 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     4021 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     4081 attacaggga cagcagagat ccactttggc cgcggctcga gggggttggg gttgcgcctt
     4141 ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa
     4201 cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg
     4261 atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg
     4321 gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact
     4381 agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct
     4441 gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg
     4501 cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca
     4561 ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac
     4621 ctctctcccc agggggatcc accggagctt accatgaccg agtacaagcc cacggtgcgc
     4681 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     4741 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     4801 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4861 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4921 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4981 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     5041 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5101 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5161 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5221 acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg
     5281 accgaaagga gcgcacgacc ccatgcatcg gtacctttaa gaccaatgac ttacaaggca
     5341 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     5401 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     5461 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     5521 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     5581 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     5641 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     5701 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5761 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     5821 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5881 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5941 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     6001 gccctatagt gagtcgtatt acgcgcgctc cccgggtacc gagctcgaat tcactggccg
     6061 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     6121 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     6181 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     6241 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     6301 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     6361 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     6421 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     6481 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6541 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6601 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6661 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6721 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6781 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6841 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6901 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6961 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7021 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7081 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7141 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7201 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7261 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7321 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7381 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7441 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7501 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7561 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7621 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7681 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7741 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7801 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7861 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7921 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7981 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8041 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8101 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8161 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8221 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8281 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8341 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8401 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     8461 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     8521 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     8581 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     8641 aacaatttca cacaggaaac agctatgac
//
