LOCUS       sequence-402480-        8541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian lentiviral vector for the expression of the 
            Sapphire-tagged 40nm GEM under the doxycycline-inducible promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8541)
  AUTHORS   Shu T et al.
  TITLE     nucGEMs probe the biophysical properties of the nucleoplasm
  JOURNAL   bioRxiv 2022
REFERENCE   2  (bases 1 to 8541)
  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: "bioRxiv 
            2022"
COMMENT     Sequence Label: TetCMV-Pfv-Sapphire
FEATURES             Location/Qualifiers
     source          1..8541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             32..665
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    712..837
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1334..1567
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1752..1796
                     /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    2059..2174
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2392..2705
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2400..2418
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2436..2454
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2471..2489
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2507..2525
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2543..2561
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2578..2596
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2614..2632
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2679..2703
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    4541..5129
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4594..4614)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5012..5023)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5336..5969
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6097..6113)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6097..6113)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6110..6132)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6121..6137
                     /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)."
     promoter        complement(join(6145..6151,6152..6169,6170..6175))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6190..6211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6328..6345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6499..7087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6579..6598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7258..8049,8050..8118))
                     /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     7881..7900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8119..8223)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8271..8405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8308..8327)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8362..8381
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tttgttaaat atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga
       61 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac
      121 tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta
      181 ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac
      241 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac
      301 agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc
      361 tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg
      421 ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg
      481 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
      541 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
      601 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
      661 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
      721 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
      781 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
      841 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
      901 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
      961 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     1021 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     1081 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     1141 gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg
     1201 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     1261 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     1321 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     1381 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     1441 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     1501 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     1561 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     1621 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     1681 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     1741 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     1801 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     1861 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     1921 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg
     1981 aaggaataga agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg
     2041 gatctcgacg gtatcgcctt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagttta tttaattaaa
     2221 gtcgaccaat tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat
     2281 tgctgcaggc gcagaactgg taggtatgga agatctctag aagctgggta ccagctgcta
     2341 gcaagcttgc tagcggccga ggcgtatcac gaggcccttt cgtcttcact cgagtttact
     2401 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgatg
     2461 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
     2521 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
     2581 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
     2641 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
     2701 atcgcgaatt cgccaccatg tctagactct ctatcaaccc gacattgata aaccgggaca
     2761 aaccgtatac taaagaagaa cttatggaaa tcctgcggct ggcaatcatt gctgagttgg
     2821 acgcgatcaa cctctatgaa cagatggcac ggtattctga agatgagaac gtaaggaaga
     2881 ttctccttga cgtcgctcgg gaggagaagg cgcacgtggg tgagtttatg gcactccttc
     2941 ttaaccttga ccctgagcaa gtgactgagt tgaaaggcgg cttcgaagag gtaaaagaac
     3001 tgacaggcat tgaagcccat attaacgata ataaaaaaga agagtccaac gtcgaatatt
     3061 ttgaaaaact ccgctccgct ctgttggatg gagtaaataa ggggaggagt ctccttaagc
     3121 atctgccagt tacacgcata gaagggcagt cctttcgagt agacattatt aaatttgagg
     3181 atggggtgcg cgtcgttaaa caggaataca agcccattcc acttctcaaa aagaaatttt
     3241 acgtaggaat tagagaactg aatgatggta cgtacgacgt ttctattgcc acgaaggcag
     3301 gtgaattgtt ggttaaggac gaggaaagcc ttgtgataag agaaatcctg tcaactgaag
     3361 ggataaagaa aatgaagttg agtagctggg acaatcccga ggaggcgctg aatgatctta
     3421 tgaatgccct tcaggaagcc tctaatgctt ccgctggacc tttcggtctg ataattaatc
     3481 caaagagata cgcgaaactt cttaaaatct atgagaaatc aggaaaaatg ttggtagaag
     3541 tgctgaaaga aatttttcgg ggtggcatca tagtcacact gaacatagat gaaaacaagg
     3601 ttattatttt tgcaaatact ccagcggtgc ttgacgtagt tgttgggcag gacgtaacgc
     3661 tccaggaatt gggacctgag ggggacgacg tggccttttt ggtgtcagaa gccatcggca
     3721 tccgaataaa gaaccccgaa gcgattgttg tcctcgaatg gtcagggtca ggttctggct
     3781 caggtagcgg gagtgggcgg ccgctgatgt ctaaggggga ggagcttttc acaggtgtag
     3841 tacctattct tgttgaattg gatggcgatg tcaatggaca caaattctcc gtctcagggg
     3901 agggagaggg agacgctacc tatggaaagt tgactctcaa attcatctgc acaacgggta
     3961 agttgccggt cccttggccg acactcgtta ctacgttcag ctatggagta cagtgtttcg
     4021 cgcgctaccc cgaccatatg aaacaacacg actttttcaa atccgcaatg cccgagggat
     4081 acgttcagga acggactatt ttctttaaag acgacggtaa ttacaaaaca cgcgccgaag
     4141 ttaagttcga aggagatact ttggttaacc gaatagagct gaaaggcatt gatttcaaag
     4201 aggacgggaa catcctgggt cataaacttg agtataattt taattcccac aacgtgtata
     4261 taatggcaga taaacagaag aatggcatca aggttaactt caagattcgg cacaatattg
     4321 aagacggtag tgtacagctg gctgaccact atcagcagaa tacgccaatt ggtgatgggc
     4381 ccgtcttgtt gccagataat cactacctct caatccagag tgcccttagt aaagatccaa
     4441 acgaaaagcg ggaccacatg gttttgttgg agtttgtcac agctgctggg attacccacg
     4501 ggatagacga gttgtataaa tgactcgacg cgtctggaac aatcaacctc tggattacaa
     4561 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     4621 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4681 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     4741 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     4801 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     4861 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     4921 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     4981 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     5041 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     5101 tcggatctcc ctttgggccg cctccccgcc tggaattaat tctgcagtcg agacctagaa
     5161 aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta
     5221 gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca
     5281 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa
     5341 gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa
     5401 ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc
     5461 tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa
     5521 ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga
     5581 gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg
     5641 catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg
     5701 gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc
     5761 atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
     5821 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     5881 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     5941 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     6001 tttataactt gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gctagcgttt
     6061 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6121 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6181 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6241 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6301 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6361 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6421 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6481 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6541 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6601 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6661 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6721 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6781 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6841 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6901 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6961 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7021 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7081 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7141 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7201 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7261 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7321 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7381 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7441 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7501 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7561 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7621 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7681 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7741 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7801 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7861 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7921 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7981 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8041 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8101 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8161 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8221 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta
     8281 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     8341 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     8401 ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta
     8461 ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat
     8521 tttcatttta aagaaattgt a
//
