LOCUS       sequence-134470-        8547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express copGFP and iCre under CB promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8547)
  TITLE     lenti vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8547)
  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            "Unpublished"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: pCDH-CB-copGFP-T2A-iCre
FEATURES             Location/Qualifiers
     source          1..8547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             234..414
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1076..1309
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1798..1915
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     intron          2567..2663
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             2765..3427
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     CDS             3512..3565
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3572..3592
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    4642..5230
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4695..4715)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5113..5124)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5304..5537
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5609..5730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5646..5665)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5700..5719
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5749..5884
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5922..5938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5922..5938)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5935..5957)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5946..5962
                     /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(5970..5976,5977..5994,5995..6000))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6015..6036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6153..6170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6324..6912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7083..7874,7875..7943))
                     /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     7706..7725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7944..8048)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8116..8134
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8172..8194)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8294..8313
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8507..8529
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8521..8538
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8522..8538
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 ttaattaagg tacctctggt cgttacataa cttacggtaa atggcccgcc tggctgaccg
     1981 cccaacgacc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     2041 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     2101 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     2161 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctact
     2221 cgaggccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     2281 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggggggc
     2341 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
     2401 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
     2461 ggcggcccta taaaaagcga agcgcgcggc gggcgggagc gggatcagcc accgcggtgg
     2521 cggccctaga gtcgatcgag gaactgaaaa accagaaagt taactggtaa gtttagtctt
     2581 tttgtctttt atttcaggtc cccgatccgg tggtggtgca aatcaaagaa ctgctcctca
     2641 gtggatgttg cctttacttc taggcctgta cggaagtgtt acttctgctc taaaagctgc
     2701 ggaattgtac ccgcggccga tccaccggtc gggatccacc atggagagcg acgagagcgg
     2761 cctgcccgcc atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga
     2821 gctggtgggc ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag
     2881 caccaaaggc gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt
     2941 ctaccacttc ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa
     3001 cggcggctac accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag
     3061 cttcagctac cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg
     3121 cttccccgag gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga
     3181 gcacctgcac cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct
     3241 gcgcgacggc ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat
     3301 ccaccccagc atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct
     3361 gcacagcaac accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccattgc
     3421 cttcgccaga tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg
     3481 ctccaccgga tctcgcaagc ttggcagtgg agagggcaga ggaagtctgc taacatgcgg
     3541 tgacgtcgag gagaatcctg gcccaatggt gcccaagaag aagaggaaag tctccaacct
     3601 gctgactgtg caccaaaacc tgcctgccct ccctgtggat gccacctctg atgaagtcag
     3661 gaagaacctg atggacatgt tcagggacag gcaggccttc tctgaacaca cctggaagat
     3721 gctcctgtct gtgtgcagat cctgggctgc ctggtgcaag ctgaacaaca ggaaatggtt
     3781 ccctgctgaa cctgaggatg tgagggacta cctcctgtac ctgcaagcca gaggcctggc
     3841 tgtgaagacc atccaacagc acctgggcca gctcaacatg ctgcacagga gatctggcct
     3901 gcctcgccct tctgactcca atgctgtgtc cctggtgatg aggagaatca gaaaggagaa
     3961 tgtggatgct ggggagagag ccaagcaggc cctggccttt gaacgcactg actttgacca
     4021 agtcagatcc ctgatggaga actctgacag atgccaggac atcaggaacc tggccttcct
     4081 gggcattgcc tacaacaccc tgctgcgcat tgccgaaatt gccagaatca gagtgaagga
     4141 catctcccgc accgatggtg ggagaatgct gatccacatt ggcaggacca agaccctggt
     4201 gtccacagct ggtgtggaga aggccctgtc cctgggggtt accaagctgg tggagagatg
     4261 gatctctgtg tctggtgtgg ctgatgaccc caacaactac ctgttctgcc gggtcagaaa
     4321 gaatggtgtg gctgcccctt ctgccacctc ccaactgtcc acccgggccc tggaagggat
     4381 ctttgaggcc acccaccgcc tgatctatgg tgccaaggat gactctgggc agagatacct
     4441 ggcctggtct ggccactctg ccagagtggg tgctgccagg gacatggcca gggctggtgt
     4501 gtccatccct gaaatcatgc aggctggtgg ctggaccaat gtgaacattg tgatgaacta
     4561 catcagaaac ctggactctg agactggggc catggtgagg ctgctcgagg atggggactg
     4621 agcggccgcg aattcgtcga caatcaacct ctggattaca aaatttgtga aagattgact
     4681 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4741 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4801 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4861 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4921 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     4981 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     5041 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5101 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5161 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5221 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     5281 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct
     5341 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     5401 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     5461 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     5521 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     5581 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     5641 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5701 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     5761 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5821 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5881 ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg
     5941 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6001 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6061 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6721 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8041 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     8101 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     8161 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     8221 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     8281 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     8341 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     8401 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     8461 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     8521 tgtaaaacga cggccagtgc caagctg
//
