LOCUS       sequence-460688-        8321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Label centrosomes with lentivirus by encoding a fusion protein 
            consisting of the photo-convertible protein Kaede fused to Centrin 1
            (CETN1), an integral component of centrosomes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8321)
  TITLE     Targeted Localization of the Mother Centrosome in CD8+ T Cells 
            Undergoing Asymmetric Cell Division Promotes Memory Formation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8321)
  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: 
            "Unpublished"
COMMENT     Sequence Label: 1993_pCCLsin.PPT.hPGK.KaedeCETN1.PRE
FEATURES             Location/Qualifiers
     source          1..8321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(131..199,200..991)
                     /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(349..368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1162..1750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1651..1670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1904..1921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2038..2059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2074..2079,2080..2097,2098..2104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2112..2128
                     /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     2117..2139
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2136..2152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2136..2152
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2171..2191
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2173..2191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        2292..2671
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2672..2875
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2826..2846
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2890..3070
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3117..3242
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3735..3968
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4153..4197
                     /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    4464..4581
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4627..5137
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     5056..5074
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      5153..5162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5159..5836
                     /codon_start=1
                     /product="Kaede green-to-red photoconvertible fluorescent 
                     protein (Ando et al., 2002)"
                     /label=Kaede
                     /translation="MVSLIKPEMKIKLLMEGNVNGHQFVIEGDGKGHPFEGKQSMDLVV
                     KEGAPLPFAYDILTTAFHYGNRVFAKYPDHIPDYFKQSFPKGFSWERSLMFEDGGVCIA
                     TNDITLKGDTFFNKVRFDGVNFPPNGPVMQKKTLKWEASTEKMYLRDGVLTGDITMALL
                     LKGDVHYRCDFRTTYKSRQEGVKLPGYHFVDHCISILRHDKDYNEVKLYEHAVAHSGLP
                     DNVK"
     misc_feature    6383..6971
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6436..6456)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6854..6865)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7058..7291
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7363..7484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7400..7419)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7454..7473
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7524..7659
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7679..7698)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7680..7698)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7708..7725)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7708..7724)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7717..7739)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7866..8294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7953..7972)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8163..8184
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2161 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttggcc
     2221 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
     2281 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2341 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2401 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2461 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2521 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2581 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2641 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2701 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2761 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2821 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
     2881 agtgaaccgg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     2941 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     3001 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     3061 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
     3121 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
     3181 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     3241 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     3301 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     3361 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     3421 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     3481 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     3541 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
     3601 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     3661 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     3721 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     3781 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     3841 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     3901 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     3961 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     4021 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     4081 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     4141 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     4201 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     4261 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     4321 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     4381 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     4441 ggatctcgac ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca
     4501 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     4561 attacaaaaa ttcaaaattt tatcgatcac gagactagcc tcgagaagct tgatatcgaa
     4621 ttccacgggg ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg
     4681 ctgctctggg cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt
     4741 cacgtccgtt cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac
     4801 gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg
     4861 acaaacggaa gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat
     4921 ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga
     4981 gagcagcggc cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct
     5041 gttcctgccc gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg
     5101 ctccctcgtt gaccgaatca ccgacctctc tccccagggg gatccaccgg tcgccaccat
     5161 ggtgagtctg attaaaccag aaatgaagat caagctgctt atggaaggca atgtaaacgg
     5221 gcaccagttt gttattgagg gagatggaaa aggccatcct tttgagggaa aacagagtat
     5281 ggaccttgta gtcaaagaag gcgcacctct cccttttgcc tacgatatct tgacaacagc
     5341 attccattat ggtaacaggg tttttgctaa atacccagac catataccag actacttcaa
     5401 gcagtcgttt cccaaagggt tttcttggga gcgaagcctg atgttcgagg acgggggcgt
     5461 ttgcatcgct acaaatgaca taacactgaa aggagacact ttttttaaca aagttcgatt
     5521 tgatggcgta aactttcccc caaatggtcc tgttatgcag aagaagactc tgaaatggga
     5581 ggcatccact gagaaaatgt atttgcgtga tggagtgttg acgggcgata ttaccatggc
     5641 tctgctgctt aaaggagatg tccattaccg atgtgacttc agaactactt acaaatctag
     5701 gcaggagggt gtcaagttgc caggatatca ctttgtcgat cactgcatca gcatattgag
     5761 gcatgacaaa gactacaacg aggttaagct gtatgagcat gctgttgccc attctggatt
     5821 gccggacaac gtcaaggtcg acggtaccat ggcttccggc ttcaagaagc ccagcgctgc
     5881 ctccaccggc caaaagagaa aggtggcacc taagcccgag ctcactgagg atcagaagca
     5941 agaagttcgg gaagcatttg acctcttcga cgtggacgga agtgggacca tcgacgcgaa
     6001 ggagctgaag gtggccatga gagcgctggg cttcgaaccc aggaaggaag agatgaagaa
     6061 aatgatctcc gaggtggaca gggaaggcac ggggaagatc agcttcaatg acttcctggc
     6121 cgtgatgacg cagaagatgt ccgagaagga caccaaagaa gaaatcctga aggccttcag
     6181 gctctttgat gacgatgaga ccgggaagat ctcgttcaaa aacctgaagc gtgtggccaa
     6241 cgagctgggg gagaacctca cggatgagga gctgcaggag atgatcgacg aagctgatcg
     6301 ggatggggac ggcgaagtga acgaggagga gttccttcgg atcatgaaga agaccagcct
     6361 ttactgaagc ggccgcgtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     6421 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     6481 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     6541 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     6601 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     6661 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     6721 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     6781 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     6841 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     6901 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     6961 cgcctccccg cctggaattc gagctcggta cctttaagac caatgactta caaggcagct
     7021 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     7081 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     7141 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     7201 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     7261 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     7321 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     7381 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     7441 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     7501 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     7561 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     7621 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     7681 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     7741 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     7801 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     7861 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     7921 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     7981 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     8041 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     8101 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     8161 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     8221 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     8281 atttaacgcg aattttaaca aaatattaac gcttacaatt t
//
