LOCUS       Exported                8800 bp DNA     circular SYN 09-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8800)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8800
                     /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    1804..1921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2011..2214
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2317..2901
                     /codon_start=1
                     /label=cAMP (human)
                     /translation="MKTQRDGHSLGRWSLVLLLLGLVMPLAIIAQVLSYKEAVLRAIDG
                     INQRSSDANLYRLLDLDPRPTMDGDPDTPKPVSFTVKETVCPRTTQQSPEDCDFKKDGL
                     VKRCMGTVTLNQARGSFDISCDKDNKRFALLGDFFRKSKEKIGKEFKRIVQRIKDFLRN
                     LVPRTESGSDYKDHDGDYKDHDIDYKDDDDK"
     CDS             2833..2898
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        2942..3153
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3166..3434
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      3463..3472
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3493..4155
                     /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             4225..4278
                     /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             4279..4878
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    4879..5467
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5350..5361)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5541..5774
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5846..5967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6007..6142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6175..6191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6199..6215
                     /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(6223..6253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6268..6289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6577..7165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7336..8196)
                     /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"
     promoter        complement(8197..8301)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8775..8791
                     /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 ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc ctgatacgaa ctcggaattc gccaccatga agacccaaag ggatggccac
     2341 tccctggggc ggtggtcact ggtgctcctg ctgctgggcc tggtgatgcc tctggccatc
     2401 attgcccagg tcctcagcta caaggaagct gtgcttcgtg ctatagatgg catcaaccag
     2461 cggtcctcgg atgctaacct ctaccgcctc ctggacctgg accccaggcc cacgatggat
     2521 ggggacccag acacgccaaa gcctgtgagc ttcacagtga aggagacagt gtgccccagg
     2581 acgacacagc agtcaccaga ggattgtgac ttcaagaagg acgggctggt gaagcggtgt
     2641 atggggacag tgaccctcaa ccaggccagg ggctcctttg acatcagttg tgataaggat
     2701 aacaagagat ttgccctgct gggtgatttc ttccggaaat ctaaagagaa gattggcaaa
     2761 gagtttaaaa gaattgtcca gagaatcaag gattttttgc ggaatcttgt acccaggaca
     2821 gagtccggat ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac
     2881 aaggatgacg atgacaagta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     2941 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     3001 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     3061 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     3121 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     3181 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     3241 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     3301 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     3361 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     3421 cgccgttaca gatccaagct gtgaccggcg cctacgctag acgccaccat ggagagcgac
     3481 gagagcggcc tgcccgccat ggagatcgag tgccgcatca ccggcaccct gaacggcgtg
     3541 gagttcgagc tggtgggcgg cggagagggc acccccaagc agggccgcat gaccaacaag
     3601 atgaagagca ccaaaggcgc cctgaccttc agcccctacc tgctgagcca cgtgatgggc
     3661 tacggcttct accacttcgg cacctacccc agcggctacg agaacccctt cctgcacgcc
     3721 atcaacaacg gcggctacac caacacccgc atcgagaagt acgaggacgg cggcgtgctg
     3781 cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt caaggtggtg
     3841 ggcaccggct tccccgagga cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc
     3901 accgtggagc acctgcaccc catgggcgat aacgtgctgg tgggcagctt cgcccgcacc
     3961 ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg acagccacat gcacttcaag
     4021 agcgccatcc accccagcat cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg
     4081 gaggagctgc acagcaacac cgagctgggc atcgtggagt accagcacgc cttcaagacc
     4141 cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga cggcaccgcc
     4201 ggacccggct ccaccggatc tcgcgagggc agaggaagtc ttctaacatg cggtgacgtg
     4261 gaggagaatc ccggccctat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4321 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4381 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     4441 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     4501 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     4561 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     4621 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     4681 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     4741 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     4801 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     4861 cgcaagcccg gtgcctgaaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     4921 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     4981 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5041 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     5101 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     5161 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     5221 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     5281 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5341 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5401 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5461 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     5521 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagat aagatctgct
     5581 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     5641 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     5701 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     5761 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     5821 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     5881 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5941 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     6001 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     6061 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     6121 tgaggaggct tttttggagg cctagacttt tgcagagacc aaattcgtaa tcatgtcata
     6181 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6241 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6301 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6361 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6421 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6481 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6541 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6601 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6661 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6721 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6781 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6841 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6901 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6961 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7021 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7081 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7141 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7201 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7261 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7321 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7381 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7441 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7501 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7561 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7621 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7681 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7741 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7801 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7861 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7921 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7981 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8041 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8101 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8161 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8221 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8281 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     8341 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     8401 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     8461 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     8521 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     8581 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     8641 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     8701 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     8761 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
