LOCUS       Exported                8778 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 8778)
  AUTHORS   vfsd
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(6987..8778,1..6986)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    236..352
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
     enhancer        409..712
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        713..916
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             1015..1722
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    1723..1788
                     /label=MCS
                     /note="multiple cloning site"
     promoter        1821..2320
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             2341..2940
                     /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    2954..3542
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     CDS             complement(3425..3436)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3749..4382
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4511..4527)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4535..4551
                     /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(4559..4589)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4604..4625
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4913..5501)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5672..6532)
                     /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(6533..6637)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6685..6819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             6987..7620
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7667..7792
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    8289..8522
                     /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 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
       61 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
      121 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
      181 gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta tcgcctttaa
      241 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
      301 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
      361 ttacagggac agcagagatc cagtttatcg ataagcttgg gagttccgcg ttacataact
      421 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      481 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      541 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      601 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      661 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      721 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      781 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      841 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      901 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
      961 ttttgacctc catagaagac accgactcta ctagaggatc taccggtcgc caccatggtg
     1021 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1081 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1141 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1201 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     1261 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1321 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1381 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1441 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     1501 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     1561 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     1621 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     1681 gagggccgcc actccaccgg cggcatggac gagctgtaca agtccggact cagatctcga
     1741 gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga
     1801 taactgatca taattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg
     1861 cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca
     1921 ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc
     1981 ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac
     2041 gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca
     2101 atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg
     2161 ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg
     2221 ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc
     2281 tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc caagcttacc
     2341 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     2401 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     2461 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     2521 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     2581 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     2641 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     2701 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     2761 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     2821 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     2881 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     2941 ccgcgtctgg aacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3001 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3061 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3121 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3181 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3241 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3301 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     3361 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3421 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3481 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3541 gcctggaatt aattctgcag tcgagaccta gaaaaacatg gagcaatcac aagtagcaat
     3601 acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt
     3661 tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     3721 agccactttt taaaagaaaa gaggggactg gaagggctaa ttcactccca acgaagacaa
     3781 gatatccttg atctgtggat ctaccacaca caaggctact tccctgatta gcagaactac
     3841 acaccagggc caggggtcag atatccactg acctttggat ggtgctacaa gctagtacca
     3901 gttgagccag ataaggtaga agaggccaat aaaggagaga acaccagctt gttacaccct
     3961 gtgagcctgc atgggatgga tgacccggag agagaagtgt tagagtggag gtttgacagc
     4021 cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa gaactgctga
     4081 tatcgagctt gctacaaggg actttccgct ggggactttc cagggaggcg tggcctgggc
     4141 gggactgggg agtggcgagc cctcagatcc tgcatataag cagctgcttt ttgcctgtac
     4201 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     4261 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     4321 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     4381 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     4441 agagagtgag aggccttgac attgctagcg ttttaccgtc gacctctagc tagagcttgg
     4501 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4561 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4621 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4681 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4741 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4801 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4861 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4921 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4981 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5041 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5101 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5161 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5221 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5281 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5341 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5401 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5461 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5521 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5581 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5641 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5701 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5761 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5821 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5881 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5941 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6001 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6061 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6121 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6181 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6241 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6301 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6361 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6421 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6481 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6541 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6601 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6661 ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa
     6721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6781 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
     6841 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
     6901 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
     6961 taaatatgta ctacaaactt agtagttgga agggctaatt cactcccaaa gaagacaaga
     7021 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac
     7081 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     7141 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt
     7201 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg
     7261 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata
     7321 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg
     7381 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg
     7441 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     7501 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     7561 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     7621 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     7681 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     7741 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     7801 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     7861 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     7921 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     7981 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     8041 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     8101 aaacaaaagt aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga
     8161 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     8221 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     8281 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     8341 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     8401 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     8461 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     8521 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     8581 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     8641 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     8701 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     8761 tttaacataa caaattgg
//
