LOCUS       sequence-472159-        7522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express D2GFP from a 0.5 bp synapsin promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7522)
  TITLE     Ploski Lab Plasmids 2025
REFERENCE   2  (bases 1 to 7522)
  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: pLenti-0.5Syn-D2GFP-WPRE
FEATURES             Location/Qualifiers
     source          1..7522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    175..296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(212..231)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     266..285
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      336..471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(491..510)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(492..510)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(520..537)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(520..536)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(529..551)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      678..1133
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(765..784)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     975..996
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1159..1263
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1264..1332,1333..2124)
                     /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(1482..1501)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2295..2883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2784..2803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3037..3054
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3171..3192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3207..3212,3213..3230,3231..3237)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3245..3261
                     /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     3250..3272
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3269..3285
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3269..3285
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3304..3324
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3306..3324
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3352..3578
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3579..3759
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3806..3931
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4424..4657
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4842..4886
                     /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    5153..5270
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      5846..5855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5852..5854,5855..5857,5858..6568,6569..6574,
                     6575..6697)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(5897..5918)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6158..6177)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6505..6526
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6724..7312
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6777..6797)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7195..7206)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             join(7392..7522,1..103)
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
ORIGIN
        1 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
       61 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
      121 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
      181 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
      241 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
      301 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      361 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      421 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
      481 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
      541 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
      601 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      661 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
      721 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
      781 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
      841 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
      901 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
      961 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     1021 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     1081 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     1141 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1201 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1261 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1321 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1381 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1441 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1501 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1561 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1621 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1681 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     1741 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     1801 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     1861 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1921 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1981 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2041 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2101 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     2161 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2221 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2281 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2341 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2401 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2461 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2521 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2581 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2641 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2701 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2761 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2821 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2881 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2941 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3001 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3061 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3121 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     3181 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     3241 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     3301 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
     3361 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     3421 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
     3481 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     3541 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     3601 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     3661 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     3721 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     3781 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     3841 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     3901 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     3961 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     4021 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     4081 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     4141 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     4201 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     4261 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     4321 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     4381 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     4441 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     4501 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     4561 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     4621 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     4681 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     4741 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     4801 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     4861 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     4921 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     4981 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     5041 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     5101 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcggtta acttttaaaa
     5161 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     5221 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt atcgatacaa
     5281 gtatctggga agggtaacaa tgcaagttag cctgaggatt taggaggagg ctgaaaaaca
     5341 gagtaggagc cttactacgg gtccagaccc tacggacaag aacccccact cccactcccc
     5401 aaattgcgca ttccctcccc catcagaggg ggaggggaag aggatgcagc gcggcgcggc
     5461 gcgtgcgcac tgtcggattt agtaccgcgg acagagcctt cgcccccgct gccggcgcgc
     5521 gccaccacct ccccagcacc aaaggcgggc tgacgtcact ctccagccct ccccaaactc
     5581 ccctacctca ccgccttggt cgcgtccgtg cagcggtgag tccagtcggg ccgcaccaca
     5641 agaggtgcaa gatagggggg tgcaggcgcg accatacgct ctgcggcggc agagcctcag
     5701 cgctgcctca gtctgcagcg ggcagcagag gagtcgcgtc gtgccagaga gcgccgccgt
     5761 gctcctgagc cccttgcgct ccgcccccgc ggcccaccga cccactgccc cttggatcca
     5821 ctagtccagt gtggtggaat tcgccgccac catggtgagc aagggcgagg agctgttcac
     5881 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     5941 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     6001 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     6061 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     6121 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     6181 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     6241 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     6301 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     6361 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     6421 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     6481 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     6541 cactctcggc atggacgagc tgtacaagaa gcttagccat ggcttcccgc cggaggtgga
     6601 ggagcaggat gatggcacgc tgcccatgtc ttgtgcccag gagagcggga tggaccgtca
     6661 ccctgcagcc tgtgcttctg ctaggatcaa tgtgtagacg cgtaccggtt agtaatgatc
     6721 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     6781 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     6841 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     6901 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     6961 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     7021 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     7081 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     7141 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     7201 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     7261 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggcgat
     7321 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     7381 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     7441 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     7501 cccactgctt aagcctcaat aa
//
