LOCUS       sequence-364561-        9239 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral vector of constitutive expression of mNeonGreen2(1-10).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9239)
  AUTHORS   Feng S, Sekine S, Pessino V, Li H, Leonetti MD, Huang B
  TITLE     Improved split fluorescent proteins for endogenous protein labeling.
  JOURNAL   Nat Commun. 2017 Aug 29;8(1):370. doi: 10.1038/s41467-017-00494-8.
  PUBMED    28851864
REFERENCE   2  (bases 1 to 9239)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2017-08-29"; volume: "8(1):370. doi"; pages: " 
            10.1038/s41467-017-00494-8"
COMMENT     Sequence Label: pSFFV_mNG2(11)1-10
FEATURES             Location/Qualifiers
     source          1..9239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             145..378
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    546..680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(570..589)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     624..643
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1105..1125)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          1255..1320
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     promoter        complement(2454..2783)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2468..2603
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2768..2788
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2858..2875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3029..3617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3109..3128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3788..4579,4580..4648))
                     /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     4411..4430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4649..4753)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4821..4839
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4877..4899)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4999..5018
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5232..5251
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5541..5559
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5541..5558
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     LTR             5584..5817
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     5907..5923
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5908..5924
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5927..6515
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6033..6044
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6442..6462
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     regulatory      6952..6961
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7291..7698)
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    7851..7968
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(8253..8297)
                     /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    8482..8715
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    join(9212..9239,1..98)
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc
       61 gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt ccctttcgct
      121 ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa aagggtctga
      181 gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga agcactcaag
      241 gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc tcccaggctc
      301 agatctggtc taaccagaga aacccagtac aagcaaaaag cagatcttgt cttcgttggg
      361 agtgaattag cccttccaac tactaagttt gtagtacata tttaacaaat acaatttctt
      421 taaaatgaaa ataattcaga ggaatcacag gtttagagta aatgaaacca caggtaattg
      481 gcagtggtaa tagggtatgg ggtgggaagt ttgggatgat tttggttagc ttgagttatc
      541 cagttgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
      601 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
      661 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
      721 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
      781 atgatctcta gtcaaggcac tatacatcaa atattcctta ttaacccctt tacaaattaa
      841 aaagctaaag gtacacaatt tttgagcata gttattaata gcagacactc tatgcctgtg
      901 tggagtaaga aaaaacagta tgttatgatt ataactgtta tgcctactta taaaggttac
      961 agaatatttt tccataattt tcttgtatag cagtgcagct ttttcctttg tggtgtaaat
     1021 agcaaagcaa gcaagagttc tattactaaa cacagcatga ctcaaaaaac ttagcaattc
     1081 tgaaggaaag tccttggggt cttctacctt tctcttcttt tttggaggag tagaatgttg
     1141 agagtcagca gtagcctcat catcactaga tggcatttct tctgagcaaa acaggttttc
     1201 ctcattaaag gcattccacc actgctccca ttcatcagtt ccataggttg gaatctaaaa
     1261 tacacaaaca attagaatca gtagtttaac acattataca cttaaaaatt ttatatttac
     1321 cttagagctt taaatctctg taggtagttt gtccaattat gtcacaccac agaagtaagg
     1381 ttccttcaca aagatccggg gcccactcat aaatccagtt gccgccacgg tagccaatca
     1441 ccgtatcgta taaatcatcg tcggtacgtt cggcatcgct catcacaata cgtgcctgga
     1501 cgtcgaggat ttcgcgtggg tcaatgccgc gccagatcca catcagacgg ttaatcatgc
     1561 gataccagtg agggatggtt ttaccatcaa gggccgactg cacaggcggt tgtgcgccgt
     1621 gattaaagcg gcggactagc gtcgaggttt caggatgttt aaagcggggt ttgaacaggg
     1681 tttcgctcag gtttgcctgt gtcatggatg cagcctccag aatacttact ggaaactatt
     1741 gtaacccgcc tgaagttaaa aagaacaacg cccggcagtg ccaggcgttg aaaagattag
     1801 cgaccggaga ttggcgggac gaatacgacg cccatatccc acggctgttc aatccaggta
     1861 tcttgcggga tatcaacaac atagtcatca accagcggac gaccagccgg ttttgcgaag
     1921 atggtgacaa agtgcgcttt tggatacatt tcacgaatcg caaccgcagt accaccggta
     1981 tccaccaggt catcaataac gatgaagcct tcgccatcgc cttctgcgcg tttcagcact
     2041 ttaagctcgc gctggttgtc gtgatcgtag ctggaaatac aaacggtatc gacatgacga
     2101 atacccagtt cacgcgccag taacgcaccc ggtacagacc gccacggctt acggcaataa
     2161 tgcctttcca ttgttcagaa ggcatcagtc ggcttgcgag tttacgtgca tggatctgca
     2221 acatgtccca ggtgacgatg tatttttcgc tcatgtgaag tgtcccagcc tgtttatcta
     2281 cggcttaaaa agtgttcgag gggaaaatag gttgcgcgag attatagaga tctggcgcac
     2341 taaaaaccag tatttcacat gagtccgcgt ctttttacgc actgcctctc cctgacgcgg
     2401 gataaagtgg tattctcaaa catatctcgc aagcctgtct tgtgtccaag ctttttgcaa
     2461 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg
     2521 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     2581 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     2641 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     2701 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     2761 tttccacacc ctaactgaca cacattccac agctgcatta atgaatcggc caacgcgcgg
     2821 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2881 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2941 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3001 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3061 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3121 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3181 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3241 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3301 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3361 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3421 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3481 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3541 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3601 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3661 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3721 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3781 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3841 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3901 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3961 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4021 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4081 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4141 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4201 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4261 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4321 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4381 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4441 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4501 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4561 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4621 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4681 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4741 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     4801 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     4861 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     4921 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     4981 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cattcgacgc
     5041 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     5101 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     5161 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     5221 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     5281 atgccggcca cgatgcgtcc ggcgtagagg atctggctag cgatgaccct gctgattggt
     5341 tcgctgacca tttccgggtg cgggacggcg ttaccagaaa ctcagaaggt tcgtccaacc
     5401 aaaccgactc tgacggcagt ttacgagaga gatgataggg tctgcttcag taagccagat
     5461 gctacacaat taggcttgta catattgtcg ttagaacgcg gctacaatta atacataacc
     5521 ttatgtatca tacacatacg atttaggtga cactatagaa tacacggaat taattctaga
     5581 tgctgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
     5641 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
     5701 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
     5761 acccagtaca agcaaaaagc agatcttgtc ttcgttggga gtgaattagc ccttccagtc
     5821 cccccttttc ttttaaaaag tggctaagat ctacagctgc cttgtaagtc attggtctta
     5881 aaggtaccga gctcgaatta attccctcga ggtcgacggt atcgatgcgg ggaggcggcc
     5941 caaagggaga tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc
     6001 ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag
     6061 gacgtcccgc gcagaatcca ggtggcaaca caggcgagca gccaaggaaa ggacgatgat
     6121 ttccccgaca acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg
     6181 gcaaggcagg cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg
     6241 gaaaggagct gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca
     6301 gtgcacacca cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac
     6361 caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa
     6421 ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc
     6481 aatctttcac aaattttgta atccagaggt tgattatcga taagcttgat atcaagcttg
     6541 catgcctgca ggtcgactct agagtcgcgg ccgcttacat ggagtgcttg agctccgtct
     6601 tacggaacac gtacatcggc tggttcttca gatagttagc cgccattggc ttggcaaagg
     6661 tgtaggtggt ccgcgcagtg ctccggtagc gtttgccatt tacagtggtg taactccact
     6721 taaaggtact gatgatggtt ttgtcgttgg ggtaagtctt cttcgacatg caccagtccg
     6781 cagcggtcag cgtgttggtc atcacaggac cgtcagcagg gaaaccagtc cccatcacct
     6841 gggcctctcc tttgatgtgg cttccctcgt aggtgtagcg gtagttaaca gtaagggagg
     6901 caccatcttc aaactgcatt gtgcgatgga cttggtatcc ggagccatct accatggcgg
     6961 cctggaaagg cgacatcccg tcagggtagg gcaggtactg atggaagcca tacccgatat
     7021 gagggaccag aatccagggg gagaactgga ggtcaccctt ggtggacttc aggtttaact
     7081 cctcataacc atcatttgga ttgccggtac cctgacccac catgtcaaag tccacaccgt
     7141 tgatggagcc aaagatgtgt aactcatgag tcgctgggag agaggccatg ttatcctcct
     7201 cacccttgct caccatggat cccgggcccg cggtaccgca agcttgatat cctgcagacg
     7261 cgtgagaatt ctcgagagct ccagatcccc cccgggcgac tcagtctgtc ggaggactgg
     7321 cgcgccgagt gaggggttgt gagctctttt atagagctcg ggaagcagaa gcgcgcgaac
     7381 agaagcgaga agcaggctga ttggttaatt caaataaggc gcagggtcat ttcaggtcct
     7441 tgggggagcc tggaaacatc tgatgggtct taagaaactg ctgagggttg ggccatatct
     7501 ggggaccatc tgttcttggc ctcgggccgg ggccgaaact gcggtgacca tctgttcttg
     7561 gccccgggcc ggggccgaaa ctgctcaccg cagatatcct gtttggccca acgttagcta
     7621 ttttcatgta cccgcccttg atctgaactt ctctattctt ggtttggtat ttttccatgc
     7681 cttgcaaaat ggcgttactg cagctagctt gccaaaccta caggtggggt ctttcattcc
     7741 cccctttttc tggagactaa ataaaatctt ttattttatc ggggctgcag gaattcgata
     7801 tcaagcttat cgatccaaac tggatctctg ctgtccctgt aataaacccg aaaattttga
     7861 atttttgtaa tttgtttttg taattcttta gtttgtatgt ctgttgctat tatgtctact
     7921 attctttccc ctgcactgta ccccccaatc cccccttttc ttttaaaatt gtggatgaat
     7981 actgccattt ggcgataccg tcgagatccg ttcactaatc gaatggatct gtctctgtct
     8041 ctctctccac cttcttcttc tattccttcg ggcctgtcgg gtcccctcgg ggttgggagg
     8101 tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca
     8161 gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac
     8221 cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat
     8281 aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag
     8341 cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt
     8401 tccagagatt tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt
     8461 ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg
     8521 attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc
     8581 tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc
     8641 tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca
     8701 aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc
     8761 ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata taattcactt
     8821 ctccaattgt ccctcatatc tcctcctcca ggtctgaaga tcagcggccg gccgcttgct
     8881 gtgcggtggt cttacttttg ttttgctctt cctctatctt gtctaaagct tccttggtgt
     8941 cttttatctc tatcctttga tgcacacaat agagggttgc tactgtatta tataatgatc
     9001 taagttcttc tgatcctgtc tgaagggatg gttgtagctg tcccagtatt tgtctacagc
     9061 cttctgatgt ttctaacagg ccaggattaa ctgcgaatcg ttctagctcc ctgcttgccc
     9121 atactatatg ttttaattta tattttttct ttccccctgg ccttaaccga attttttccc
     9181 atcgcgatct aattctcccc cgcttaatac tgacgctctc gcacccatct ctctccttc
//
