LOCUS       Exported                8172 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 8172)
  AUTHORS   hfytfiu
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(6340..8172,1..6339)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    195..311
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
     enhancer        368..671
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        672..875
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    1010..1582
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             1584..2294
                     /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    2308..2896
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     CDS             complement(2779..2790)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3103..3736
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(3864..3880)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3888..3904
                     /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(3912..3942)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3957..3978
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4266..4854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5025..5885)
                     /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(5886..5990)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6038..6172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             6340..6973
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7020..7145
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7642..7875
                     /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 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
       61 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
      121 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
      181 tcgacggtat cgcctttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
      241 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
      301 ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttatcga taagcttggg
      361 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      421 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      481 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      541 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      601 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      661 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      721 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      781 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      841 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
      901 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgactctac tagaggatct
      961 atttccggtg aattcctcga gactagttct agagcggccg cggatcccgc ccctctctct
     1021 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     1081 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     1141 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     1201 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     1261 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     1321 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     1381 ggatagttgt ggaaagagtc aaatggctca cctcaagcgt attcaacaag gggctgaagg
     1441 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     1501 catgtgttta gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     1561 cctttgaaaa acacgatgat aatatggtga gcaagggcga ggaggataac atggccatca
     1621 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     1681 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     1741 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     1801 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1861 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1921 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1981 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     2041 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     2101 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     2161 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     2221 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     2281 agctgtacaa gtgaacgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga
     2341 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2401 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     2461 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     2521 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     2581 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     2641 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     2701 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     2761 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     2821 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     2881 tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa
     2941 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     3001 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     3061 cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact
     3121 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     3181 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct
     3241 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca
     3301 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt
     3361 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact
     3421 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga
     3481 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
     3541 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     3601 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     3661 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     3721 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     3781 aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgtttacc gtcgacctct
     3841 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3901 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3961 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4021 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4081 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4141 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4201 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4261 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4321 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4381 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4441 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4501 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4561 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4621 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4681 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4741 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4801 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4861 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4921 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4981 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5041 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5101 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5161 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5221 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5281 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5341 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5401 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5461 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5521 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5581 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5641 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5701 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5761 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5821 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5881 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5941 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6001 cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac ttgtttattg cagcttataa
     6061 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6121 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat
     6181 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa
     6241 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag
     6301 aaattgtatt tgttaaatat gtactacaaa cttagtagtt ggaagggcta attcactccc
     6361 aaagaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     6421 agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca
     6481 agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct
     6541 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga
     6601 ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca
     6661 agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc
     6721 gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt
     6781 tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6841 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6901 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6961 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     7021 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     7081 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     7141 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     7201 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     7261 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     7321 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     7381 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     7441 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccggc cgctgatctt
     7501 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     7561 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     7621 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     7681 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     7741 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     7801 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     7861 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     7921 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     7981 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     8041 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     8101 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     8161 agtaggaggc tt
//
