LOCUS       sequence-401797-        8637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of Flag-Tagged human HEXIM1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8637)
  TITLE     Alarcon lab plasmids
REFERENCE   2  (bases 1 to 8637)
  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:  pLenti-C-Myc-DDK-IRES-Puro HEXIM1
FEATURES             Location/Qualifiers
     source          1..8637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(77..736)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     651..670
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(737..841)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(916..935)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1106..1485
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1487..1685
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             1703..1883
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1930..2055
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2552..2785
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2970..3014
                     /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    3312..3429
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3512..3891
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3892..4095
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4121..4140
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4121..4139
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5292..5321
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5340..5363
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    5397..5970
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5420..5972
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5564..5581)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5791..5810
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5972..6571
                     /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(5972..5991)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6468..6488
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(6601..6634)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6690..7278
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6743..6763)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7161..7172)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7281..7297)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7282..7298)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7807..7987
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(8049..8637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8129..8148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgattac gccccgccct gccactcatc gcagtactgt tgtaattcat
      121 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
      181 gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga
      241 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
      301 agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
      361 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
      421 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
      481 cccatatcac cagctcaccg tctttcattg ccatacggaa ctccggatga gcattcatca
      541 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
      601 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact
      661 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
      721 tgattttttt ctccatactc ttcctttttc aatattattg aagcatttat cagggttatt
      781 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
      841 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
      901 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
      961 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     1021 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     1081 cgatgtacgg gccagatatc gcgttgacat tgattattga ctagttatta atagtaatca
     1141 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     1201 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     1261 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     1321 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     1381 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     1441 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     1501 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     1561 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     1621 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     1681 agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     1741 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     1801 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     1861 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     1921 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     1981 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     2041 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     2101 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     2161 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     2221 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     2281 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     2341 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccggc
     2401 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     2461 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     2521 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     2581 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     2641 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     2701 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     2761 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     2821 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     2881 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     2941 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     3001 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     3061 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     3121 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     3181 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     3241 ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta
     3301 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     3361 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     3421 caaaattttc gggtttatta cagggacagc agagatccag tttggttagt accgggcccg
     3481 ctctagacat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag
     3541 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     3601 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     3661 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     3721 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
     3781 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
     3841 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     3901 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     3961 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     4021 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     4081 tatataagca gagctcgttt agtgaaccgt cagaattttg taatacgact cactataggg
     4141 cggccgggaa ttcgtcgact ggatccggta ccgaggagat ctgccgccgc gatcgcatgg
     4201 ccgagccatt cttgtcagaa tatcaacacc agcctcaaac tagcaactgt acaggtgctg
     4261 ctgctgtcca ggaagagctg aaccctgagc gccccccagg cgcggaggag cgggtgcccg
     4321 aggaggacag taggtggcaa tcgagagcgt tcccccagtt gggtggccgt ccggggccgg
     4381 agggggaagg gagcctggaa tcccaaccac ctcccttgca gacccaggcc tgtccagaat
     4441 ctagctgcct gagagagggc gagaagggcc agaatgggga cgactcgtcc gctggcggcg
     4501 acttcccgcc gccggcagaa gtggaaccga cgcccgaggc cgagctgctc gcccagcctt
     4561 gtcatgactc cgaggccagt aagttggggg ctcctgccgc agggggcgaa gaggagtggg
     4621 gacagcagca gagacagctg gggaagaaaa aacataggag acgcccgtcc aagaagaagc
     4681 ggcattggaa accgtactac aagctgacct gggaagagaa gaaaaagttc gacgagaaac
     4741 agagccttcg agcttcaagg atccgagccg agatgttcgc caagggccag ccggtcgcgc
     4801 cctataacac cacgcagttc ctcatggatg atcacgacca ggaggagccg gatctcaaaa
     4861 ccggcctgta ctccaagcgg gccgccgcca aatccgacga caccagcgat gacgacttca
     4921 tggaagaagg gggtgaggag gatgggggca gcgatgggat gggaggggac ggcagcgagt
     4981 ttctgcagcg ggacttctcg gagacgtacg agcggtacca cacggagagc ctgcagaaca
     5041 tgagcaagca ggagctcatc aaggagtacc tggaactgga gaagtgcctc tcgcgcatgg
     5101 aggacgagaa caaccggctg cggctggaga gcaagcggct gggtggcgac gacgcgcgtg
     5161 tgcgggagct ggagctggag ctggaccggc tgcgcgccga gaacctccag ctgctgaccg
     5221 agaacgaact gcaccggcag caggagcgag cgccgctttc caagtttgga gacacgcgta
     5281 cgcggccgct cgagcagaaa ctcatctcag aagaggatct ggcagcaaat gatatcctgg
     5341 attacaagga tgacgacgat aaggtttaaa cctaggcgta gcggccgcaa attccgcccc
     5401 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     5461 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     5521 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     5581 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     5641 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     5701 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     5761 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     5821 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     5881 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     5941 gtggttttcc tttgaaaaac acgatgataa tatgaccgag tacaagccca cggtgcgcct
     6001 cgccacccgc gacgacgtcc cccgggcagt acgcaccctc gccgccgcgt tcgccgacta
     6061 ccccgccacg cgccacaccg tcgatccaga ccgccacatc gagcgggtca ccgagctgca
     6121 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     6181 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     6241 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     6301 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     6361 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     6421 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     6481 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     6541 ctggtgcatg acccgcaagc ccggtgcctg atgtacaagt aggattcgtc gagggaccta
     6601 ataacttcgt atagcataca ttatacgaag ttatacatgt ttaagggttc cggttccact
     6661 aggtacaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     6721 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     6781 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     6841 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     6901 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     6961 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     7021 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     7081 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     7141 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     7201 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     7261 tttgggccgc ctccccgcat cgataccgtc gacctcgatc gagacctaga aaaacatgga
     7321 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa
     7381 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac
     7441 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     7501 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
     7561 cctgattggc agaactacac accagggcca gggatcagat atccactgac ctttggatgg
     7621 tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac
     7681 acccgcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta
     7741 gagtggaggt ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac
     7801 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     7861 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     7921 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     7981 tctagcagca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     8041 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     8101 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     8161 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     8221 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     8281 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     8341 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     8401 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     8461 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     8521 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     8581 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaa
//
