LOCUS       Exported                8787 bp DNA     circular SYN 10-SEP-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8787)
  AUTHORS   vfsd
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(6982..8787,1..6981)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    222..338
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
     enhancer        395..698
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        699..902
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    987..1067
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1069..1788
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     promoter        1816..2315
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             2336..2935
                     /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    2949..3537
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     CDS             complement(3420..3431)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3744..4377
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4506..4522)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4530..4546
                     /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(4554..4584)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4599..4620
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4908..5496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5667..6527)
                     /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(6528..6632)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6680..6814
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             6982..7615
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7662..7787
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    8284..8517
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
       61 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
      121 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
      181 cagatccatt cgattagtga acggatctcg acggtatcgc ctttaaaaga aaagggggga
      241 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      301 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      361 gagatccagt ttatcgataa gcttgggagt tccgcgttac ataacttacg gtaaatggcc
      421 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      481 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      541 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      601 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      661 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      721 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      781 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      841 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      901 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
      961 gaagacaccg actctactag aggatcgcta gcgctaccgg actcagatct cgagctcaag
     1021 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggtcat ggtgagcaag
     1081 ggcgccgagc tgttcaccgg catcgtgccc atcctgatcg agctgaatgg cgatgtgaat
     1141 ggccacaagt tcagcgtgag cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1201 ctgaagttca tctgcaccac cggcaagctg cctgtgccct ggcccaccct ggtgaccacc
     1261 ctgagctacg gcgtgcagtg cttctcacgc taccccgatc acatgaagca gcacgacttc
     1321 ttcaagagcg ccatgcctga gggctacatc caggagcgca ccatcttctt cgaggatgac
     1381 ggcaactaca agtcgcgcgc cgaggtgaag ttcgagggcg ataccctggt gaatcgcatc
     1441 gagctgaccg gcaccgattt caaggaggat ggcaacatcc tgggcaataa gatggagtac
     1501 aactacaacg cccacaatgt gtacatcatg accgacaagg ccaagaatgg catcaaggtg
     1561 aacttcaaga tccgccacaa catcgaggat ggcagcgtgc agctggccga ccactaccag
     1621 cagaataccc ccatcggcga tggccctgtg ctgctgcccg ataaccacta cctgtccacc
     1681 cagagcgccc tgtccaagga ccccaacgag aagcgcgatc acatgatcta cttcggcttc
     1741 gtgaccgccg ccgccatcac ccacggcatg gatgagctgt acaagtgagc ggccgcgact
     1801 ctagataatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt
     1861 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca
     1921 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta
     1981 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac
     2041 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga
     2101 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc
     2161 agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg
     2221 gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg
     2281 cgctgttctc ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac
     2341 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     2401 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     2461 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     2521 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     2581 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     2641 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     2701 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     2761 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     2821 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     2881 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaccgcg
     2941 tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3001 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3061 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3121 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3181 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3241 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3301 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     3361 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3421 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3481 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     3541 gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     3601 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     3661 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     3721 ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat
     3781 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc
     3841 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
     3901 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag
     3961 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct
     4021 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg
     4081 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac
     4141 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt
     4201 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     4261 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     4321 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     4381 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     4441 gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct ctagctagag cttggcgtaa
     4501 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4561 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4621 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4681 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4741 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4801 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4861 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4921 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4981 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5041 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5101 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5161 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5221 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5281 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5341 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5401 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5461 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5521 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5581 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5641 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5701 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5761 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5821 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5881 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5941 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6001 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6061 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6121 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6181 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6241 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6301 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6361 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6421 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6481 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6541 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6601 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6661 gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6721 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6781 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa
     6841 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt
     6901 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat
     6961 atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc
     7021 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag
     7081 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
     7141 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc
     7201 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag
     7261 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag
     7321 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg
     7381 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct
     7441 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     7501 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     7561 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     7621 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     7681 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     7741 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     7801 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     7861 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     7921 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     7981 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     8041 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     8101 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     8161 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     8221 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     8281 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     8341 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     8401 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     8461 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     8521 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     8581 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     8641 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     8701 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     8761 cataacaaat tggctgtggt atataaa
//
