LOCUS       sequence-309611-        9642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector of constitutive expression of mNeonGreen3K(1-10).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9642)
  AUTHORS   Zhou S, Feng S, Brown D, Huang B
  TITLE     Improved yellow-green split fluorescent proteins for protein 
            labeling and signal amplification.
  JOURNAL   PLoS One. 2020 Nov 23;15(11):e0242592. doi: 
            10.1371/journal.pone.0242592. eCollection 2020.
  PUBMED    33227014
REFERENCE   2  (bases 1 to 9642)
  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: "PLoS One.";
            date: "2020-11-23"; volume: "15(11):e0242592. doi"; pages: " 
            10.1371/journal.pone.0242592. eCollection 2020"
COMMENT     Sequence Label: pSFFV_mNG3K(1-10)
FEATURES             Location/Qualifiers
     source          1..9642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2050..2167
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2320..2727
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3057..3066
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3506..4094
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3559..3579)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3977..3988)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4097..4113)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4098..4114)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4204..4437
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(4462..4480)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4463..4480)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4770..4789)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5003..5022)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5122..5144
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5182..5200)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5268..5372
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5373..5441,5442..6233)
                     /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(5591..5610)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6404..6992
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6893..6912
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7146..7163
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(7233..7253)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7238..7567
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7418..7553
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          8701..8766
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8896..8916
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    9341..9475
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9378..9397)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9432..9451
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccaaa tggcagtatt
     2041 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     2101 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     2161 aaattttcgg gtttattaca gggacagcag agatccagtt tggatcgata agcttgatat
     2221 cgaattcctg cagccccgat aaaataaaag attttattta gtctccagaa aaagggggga
     2281 atgaaagacc ccacctgtag gtttggcaag ctagctgcag taacgccatt ttgcaaggca
     2341 tggaaaaata ccaaaccaag aatagagaag ttcagatcaa gggcgggtac atgaaaatag
     2401 ctaacgttgg gccaaacagg atatctgcgg tgagcagttt cggccccggc ccggggccaa
     2461 gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc aagaacagat ggtccccaga
     2521 tatggcccaa ccctcagcag tttcttaaga cccatcagat gtttccaggc tcccccaagg
     2581 acctgaaatg accctgcgcc ttatttgaat taaccaatca gcctgcttct cgcttctgtt
     2641 cgcgcgcttc tgcttcccga gctctataaa agagctcaca acccctcact cggcgcgcca
     2701 gtcctccgac agactgagtc gcccgggggg gatctggagc tctcgagaat tctcacgcgt
     2761 ctgcaggata tcaagcttgc ggtaccgcgg gcccgggatc catggtgagc aagggtgagg
     2821 aggataacat ggcctctctc ccagcgactc atgagttaca catctttggc tccatcaacg
     2881 atgtggactt tgacatggtg ggtcagggta ccggcaatcc aaatgaaggt tatgaggagt
     2941 taaacctgaa gtccaccaag ggcgacctcc agttctcccc ctggattctg gtccctcata
     3001 tcgggtatgg cttccatcag tacctgccct accctgacgg gatgtcgcct ttccaggccg
     3061 ccatggtaga tggctccgga taccaagtcc atcgcacaat gcagtttgaa gatggcgcct
     3121 cccttactgt taactaccgc tacacctacg agggaagcca catcaaagga gaggcccagg
     3181 tgatagggac tggtttccct gctgacggtc ctgtgatgac caacacgctg accgctgcgg
     3241 actggtgcat gtcgaagatg acttacccca acgacaaaac catcatcagt acctttaagt
     3301 ggagttacat cactgtaaat ggcaaacgct accggagcac tgcgcggacc acctacacct
     3361 ttgccaagcc aatggcggct aactatctga agaaccagcc gatgtacgtg ttccgtaaga
     3421 cggagctcaa gcactccatg taataagcgg ccgcgactct agagtcgacc tgcaggcatg
     3481 caagcttgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     3541 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3601 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3661 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3721 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3781 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3841 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3901 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3961 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     4021 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4081 ggccgcctcc ccgcatcgat accgtcgacc tcgagggaat taattcgagc tcggtacctt
     4141 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     4201 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     4261 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     4321 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     4381 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagca
     4441 tctagaatta attccgtgta ttctatagtg tcacctaaat cgtatgtgta tgatacataa
     4501 ggttatgtat taattgtagc cgcgttctaa cgacaatatg tacaagccta attgtgtagc
     4561 atctggctta ctgaagcaga ccctatcatc tctctcgtaa actgccgtca gagtcggttt
     4621 ggttggacga accttctgag tttctggtaa cgccgtcccg cacccggaaa tggtcagcga
     4681 accaatcagc agggtcatcg ctagccagat cctctacgcc ggacgcatcg tggccggcat
     4741 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
     4801 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
     4861 cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt
     4921 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
     4981 gcgtcgaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     5041 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5101 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5161 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5221 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5281 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5341 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5401 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5461 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5521 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5581 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5641 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5701 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5761 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5821 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5881 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5941 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6001 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6061 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6121 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6181 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6241 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6301 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6361 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6421 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6481 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6541 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6601 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6661 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6721 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6781 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6841 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6901 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6961 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7021 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7081 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7141 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     7201 ccgcgcgttg gccgattcat taatgcagct gtggaatgtg tgtcagttag ggtgtggaaa
     7261 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     7321 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     7381 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     7441 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     7501 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     7561 ttgcaaaaag cttggacaca agacaggctt gcgagatatg tttgagaata ccactttatc
     7621 ccgcgtcagg gagaggcagt gcgtaaaaag acgcggactc atgtgaaata ctggttttta
     7681 gtgcgccaga tctctataat ctcgcgcaac ctattttccc ctcgaacact ttttaagccg
     7741 tagataaaca ggctgggaca cttcacatga gcgaaaaata catcgtcacc tgggacatgt
     7801 tgcagatcca tgcacgtaaa ctcgcaagcc gactgatgcc ttctgaacaa tggaaaggca
     7861 ttattgccgt aagccgtggc ggtctgtacc gggtgcgtta ctggcgcgtg aactgggtat
     7921 tcgtcatgtc gataccgttt gtatttccag ctacgatcac gacaaccagc gcgagcttaa
     7981 agtgctgaaa cgcgcagaag gcgatggcga aggcttcatc gttattgatg acctggtgga
     8041 taccggtggt actgcggttg cgattcgtga aatgtatcca aaagcgcact ttgtcaccat
     8101 cttcgcaaaa ccggctggtc gtccgctggt tgatgactat gttgttgata tcccgcaaga
     8161 tacctggatt gaacagccgt gggatatggg cgtcgtattc gtcccgccaa tctccggtcg
     8221 ctaatctttt caacgcctgg cactgccggg cgttgttctt tttaacttca ggcgggttac
     8281 aatagtttcc agtaagtatt ctggaggctg catccatgac acaggcaaac ctgagcgaaa
     8341 ccctgttcaa accccgcttt aaacatcctg aaacctcgac gctagtccgc cgctttaatc
     8401 acggcgcaca accgcctgtg cagtcggccc ttgatggtaa aaccatccct cactggtatc
     8461 gcatgattaa ccgtctgatg tggatctggc gcggcattga cccacgcgaa atcctcgacg
     8521 tccaggcacg tattgtgatg agcgatgccg aacgtaccga cgatgattta tacgatacgg
     8581 tgattggcta ccgtggcggc aactggattt atgagtgggc cccggatctt tgtgaaggaa
     8641 ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta aagctctaag
     8701 gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat tgtttgtgta
     8761 ttttagattc caacctatgg aactgatgaa tgggagcagt ggtggaatgc ctttaatgag
     8821 gaaaacctgt tttgctcaga agaaatgcca tctagtgatg atgaggctac tgctgactct
     8881 caacattcta ctcctccaaa aaagaagaga aaggtagaag accccaagga ctttccttca
     8941 gaattgctaa gttttttgag tcatgctgtg tttagtaata gaactcttgc ttgctttgct
     9001 atttacacca caaaggaaaa agctgcactg ctatacaaga aaattatgga aaaatattct
     9061 gtaaccttta taagtaggca taacagttat aatcataaca tactgttttt tcttactcca
     9121 cacaggcata gagtgtctgc tattaataac tatgctcaaa aattgtgtac ctttagcttt
     9181 ttaatttgta aaggggttaa taaggaatat ttgatgtata gtgccttgac tagagatcat
     9241 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     9301 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     9361 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     9421 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg
     9481 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc
     9541 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta
     9601 aagaaattgt atttgttaaa tatgtactac aaacttagta gt
//
