LOCUS       Exported                9004 bp ds-DNA     circular SYN 27-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9004)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Oct 27, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9004
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    2..590
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(473..484)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        604..1103
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             1124..1723
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             1861..2094
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2166..2287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2327..2462
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(2483..2501)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2511..2527)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2669..3124
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3150..3254
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3255..4115
                     /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"
     rep_origin      4286..4874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5162..5183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5198..5228
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5236..5252
                     /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     5260..5276
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5297..5315
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5343..5569
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5570..5750
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5797..5922
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6415..6648
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6833..6877
                     /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    7144..7260
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        7283..7586
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7587..7790
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      7880..7889
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             7886..8602
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             7886..8602
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    8603..8998
                     /label=ARPP19 long
ORIGIN
        1 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
       61 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
      121 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
      181 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
      241 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
      301 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
      361 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
      421 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
      481 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
      541 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattct
      601 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc
      661 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
      721 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
      781 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
      841 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
      901 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
      961 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
     1021 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     1081 cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc
     1141 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     1201 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     1261 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     1321 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     1381 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     1441 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     1501 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     1561 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     1621 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     1681 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
     1741 gcagcgcccg accgaaagga gcgcacgacc ccatgcatct cgagggcccg gtacctttaa
     1801 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     1861 tggaagggct agctcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     1921 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     1981 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     2041 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     2101 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     2161 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2221 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2281 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     2341 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2401 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2461 cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     2521 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2581 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2641 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     2701 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2761 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2821 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2881 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2941 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3001 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3061 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3121 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3181 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3241 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3301 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3361 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3421 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3481 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3541 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3601 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3661 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3721 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3781 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3841 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3901 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3961 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4021 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4081 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4141 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4201 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4261 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4321 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4381 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4441 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4501 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4561 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4621 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4681 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4741 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4801 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4861 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4921 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4981 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5041 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5101 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5161 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5221 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5281 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt
     5341 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
     5401 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
     5461 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
     5521 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
     5581 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     5641 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     5701 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
     5761 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
     5821 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
     5881 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
     5941 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
     6001 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
     6061 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     6121 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     6181 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     6241 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
     6301 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     6361 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     6421 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     6481 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     6541 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     6601 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     6661 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     6721 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     6781 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     6841 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     6901 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     6961 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     7021 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     7081 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     7141 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     7201 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     7261 atcgataagc ttgggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     7321 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     7381 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     7441 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     7501 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     7561 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     7621 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     7681 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     7741 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     7801 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccgac
     7861 tctagaggat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     7921 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     7981 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     8041 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     8101 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     8161 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     8221 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     8281 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     8341 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     8401 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     8461 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     8521 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     8581 cggcatggac gagctgtaca agatgtctgc ggaagtcccc gaggcagcct ccgcggagga
     8641 gcagaagagt gaacacaaca tgctcccttg gtctctgcaa ccatcgattc ctaacagcct
     8701 agaggaaatg gaagataaag tgactagtcc agagaaagca gaagaagcaa aattaaaagc
     8761 aagatatcct catctgggac aaaagcctgg aggttcagat ttcttaagga aacggttgca
     8821 gaaagggcaa aaatattttg attctgggga ttacaacatg gctaaagcaa aaatgaagaa
     8881 caagcaactt cctactgcag ctccggataa gacggaggtc actggtgacc acattcccac
     8941 tccgcaagac cttcctcaac ggaagccgtc ccttgttgct agcaagctgg ctggctgagt
     9001 cgac
//
