LOCUS       sequence-420159-        8471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus vector to express mNeon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8471)
  AUTHORS   Najia MA, Borrajo J, Le A, Tsai F, Huang JY, Griffith LG, Daley GQ, 
            Blainey PC
  TITLE     Live-cell transcriptomics with engineered virus-like particles
  JOURNAL   bioRxiv 2024.10.01.616098
REFERENCE   2  (bases 1 to 8471)
  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 
            2024.10.01.616098"
COMMENT     Sequence Label: pLV_EF1a_mNeon
FEATURES             Location/Qualifiers
     source          1..8471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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    1720..1837
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1976..3154
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2207..3145
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3102..3122
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3161..3170
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3419..3428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3881..3937
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3938..4312
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     misc_feature    4328..4916
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4381..4401)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4799..4810)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4988..5221
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5299..5420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5336..5355)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5390..5409
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5460..5595
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5644..5661)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5644..5660)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5653..5675)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5802..6257
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5889..5908)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6099..6120
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6283..6387
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6388..6456,6457..7248)
                     /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(6606..6625)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7419..8007
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7908..7927
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8161..8178
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8295..8316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8331..8336,8337..8354,8355..8361)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8369..8385
                     /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     8374..8396
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8393..8409
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8393..8409
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8428..8448
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8430..8448
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaatc ttgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     1741 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     1801 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     1861 agatccactt tggcgccggc tcgagggggc ccgggtgcaa agatggataa agttttaaac
     1921 agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc
     1981 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     2041 ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     2101 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     2161 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg
     2221 tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc
     2281 cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
     2341 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
     2401 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
     2461 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
     2521 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
     2581 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
     2641 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
     2701 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     2761 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     2821 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     2881 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     2941 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     3001 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     3061 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     3121 tcaaagtttt tttcttccat ttcaggtgtc gtgacgtacg gccaccatgg tcagcaaggg
     3181 cgaggaggac aacgcctctc tcccagcgac acacgagtta cacatctttg gctccatcaa
     3241 cggtgtggac ttcgacctcg tgggtcaggg caccggcaat ccaaacgacg gttacgagga
     3301 gctcaacctc aagtccacca agggcgacct ccagttctcc ccctggattc tggtccctca
     3361 tatcgggtac ggcttccatc agtacctgcc ctacccagac gggatgtcgc ctttccaggc
     3421 cgccatggtg gacggctccg gataccaagt ccatcgcaca atgcagttcg aagacggtgc
     3481 ctcccttact gtcaactacc gctacaccta cgagggaagc cacatcaaag gagaggccca
     3541 ggtcaagggg actggtttcc ctgccgacgg tcctgtcctc accaactcgc tcaccgctgc
     3601 ggactggtgc aggtcgaaga agacttaccc caacgacaaa accatcatca gtaccttcaa
     3661 gtggagttac accactggaa acggcaagcg ctaccggagc actgcgcgga ccacctacac
     3721 ctttgccaag ccaatggcgg ccaactatct caagaaccag ccgatgtacg tgttccgcaa
     3781 gacggagctc aagcactcca agaccgagct caacttcaag gagtggcaaa aggcctttac
     3841 cgacgtcatg ggcatggacg agctctacaa gggatccggc gcaacaaact tctctctgct
     3901 caaacaagcc ggcgacgtcg aagagaatcc tgggccgatg gccaagttga ccagtgccgt
     3961 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     4021 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     4081 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     4141 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     4201 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     4261 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gaacgcgtta
     4321 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4381 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4441 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4501 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4561 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4621 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4681 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     4741 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4801 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4861 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg
     4921 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     4981 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     5041 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5101 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5161 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5221 agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     5281 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5341 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5401 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     5461 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5521 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5581 gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcat attacgcgcg
     5641 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     5701 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     5761 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     5821 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5881 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5941 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6001 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6061 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     6121 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     6181 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     6241 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6301 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6361 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6421 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6481 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6541 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6601 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6661 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6721 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6781 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6841 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6901 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6961 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     7021 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     7081 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     7141 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7201 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7261 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7321 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7381 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7441 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7501 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7561 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7621 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7681 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7741 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7801 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7861 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7921 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7981 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     8041 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     8101 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     8161 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     8221 cgttggccga ttcattaatg cagctgacac gacaggtttc ccgactggaa agcgggcagt
     8281 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     8341 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     8401 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
     8461 gagctgcaag c
//
