LOCUS       sequence-12850-e        8602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8602)
  TITLE     Lentivectors expressing localized living colors
REFERENCE   2  (bases 1 to 8602)
  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: pCLX-UBI-VenusN
FEATURES             Location/Qualifiers
     source          1..8602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             385..642
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    689..814
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1306..1423
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    1783..2016
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2201..2245
                     /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"
     protein_bind    2522..2542
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        2560..3770
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3607..3626
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    3777..3801
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3807..3809,3810..3812,3813..4520)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(3852..3873)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4113..4132)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4460..4481
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4524..4544
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(4582..4606)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4624..5212
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4677..4697)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5095..5106)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(5355..5388)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             5408..5588
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5626..5645)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5691..5746
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5745..5764)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6105..6121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6105..6121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6118..6140)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6129..6145
                     /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(6153..6159,6160..6177,6178..6183))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6198..6219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6278..6473
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6324..6459
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6479..6613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6516..6535)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6570..6589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6681..6698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6852..7440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6932..6951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7611..8402,8403..8471))
                     /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     8234..8253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8472..8576)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggaggcg tggcctgggc gggactgggg agtggcgagc
      421 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
      481 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      541 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      601 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      661 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      721 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      781 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
      841 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
      901 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
      961 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1021 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1081 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaaaaa
     1141 agcacagcaa gcagcaggat cttcagacct ggaaattccc tacaatcccc aaagtcaagg
     1201 agtagtagaa tctatgaata aagaattaaa gaaaattata ggacaggtaa gagatcaggc
     1261 tgaacatctt aagacagcag tacaaatggc agtattcatc cacaatttta aaagaaaagg
     1321 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     1381 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     1441 cagcagaaat ccactttgga aaggaccagc aaagctcctc tggaaaggtg aaggggcagt
     1501 agtaatacaa gataatagtg acataaaagt agtgccaaga agaaaagcaa agatcattag
     1561 ggattatgga aaacagatgg caggtgatga ttgtgtggca agtagacagg atgaggatta
     1621 gaacatggaa aagtttagta aaacaccata aggaggagat atgagggaca attggagaag
     1681 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1741 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1801 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1861 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1921 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1981 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     2041 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     2101 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     2161 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     2221 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     2281 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     2341 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     2401 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     2461 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgctagt gattatcgga
     2521 tcaactttgt atagaaaagt tgttaattaa gggtgcagcg gcctccgcgc cgggttttgg
     2581 cgcctcccgc gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca
     2641 ggagcgttcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact
     2701 cggccttaga accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta
     2761 gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg
     2821 attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg
     2881 gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc
     2941 gctgtgatcg tcacttggtg agttgcgggc tgctgggctg gccggggctt tcgtggccgc
     3001 cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc
     3061 gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc acaaaatggc ggctgttccc
     3121 gagtcttgaa tggaagacgc ttgtaaggcg ggctgtgagg tcgttgaaac aaggtggggg
     3181 gcatggtggg cggcaagaac ccaaggtctt gaggccttcg ctaatgcggg aaagctctta
     3241 ttcgggtgag atgggctggg gcaccatctg gggaccctga cgtgaagttt gtcactgact
     3301 ggagaactcg ggtttgtcgt ctggttgcgg gggcggcagt tatgcggtgc cgttgggcag
     3361 tgcacccgta cctttgggag cgcgcgcctc gtcgtgtcgt gacgtcaccc gttctgttgg
     3421 cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca
     3481 ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt
     3541 gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta
     3601 agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag
     3661 ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga
     3721 ctagtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac aggatcccaa
     3781 ctttgtacaa aaaagcaggc tccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     3841 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3901 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg
     3961 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct
     4021 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     4081 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     4141 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     4201 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     4261 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     4321 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     4381 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc
     4441 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     4501 tcggcatgga cgagctgtac cctccaaaaa agaagagaaa ggtataagac ccgcggtgac
     4561 taactagctc gagatatcta gacccagctt tcttgtacaa agtggtgata atcgaattcc
     4621 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     4681 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     4741 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     4801 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     4861 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     4921 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     4981 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     5041 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     5101 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     5161 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgggaa
     5221 ttcccgcggt tcgctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     5281 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatgggatc
     5341 aattcaccat gggaataact tcgtatagca tacattatac gaagttatgc tgctttttgc
     5401 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     5461 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     5521 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     5581 ctctagcagc atctagttga atcgccagtg tgtcgagcta gcagatcttt ttccctctgc
     5641 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     5701 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     5761 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     5821 tgccatatgc tggctgccat gaacaaaggt ggctataaag aggtcatcag tatatgaaac
     5881 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     5941 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     6001 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     6061 tatgaagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     6121 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6181 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6241 gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca
     6301 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     6361 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     6421 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa
     6481 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6541 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6601 tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6661 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6721 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6781 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6841 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6901 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6961 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7021 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     7081 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7141 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7201 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7261 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7321 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7381 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7441 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7501 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7561 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7621 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7681 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7741 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7801 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7861 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7921 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7981 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8041 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8101 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8161 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8221 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8281 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8341 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8401 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8461 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8521 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8581 tttccccgaa aagtgccacc tg
//
