LOCUS       Exported                9137 bp ds-DNA     circular SYN 28-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9137)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Aug 29, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9137
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2636..3083
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3090..3099
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3096..3761
                     /codon_start=1
                     /product="monomeric Keima-Red fluorescent protein, also 
                     known as mKeima"
                     /label=mKeima-Red
                     /translation="MVSVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVKLTV
                     TKGGPLPFAWDILSPQLQYGSIPFTKYPEDIPDYFKQSFPEGYTWERSMNFEDGAVCTV
                     SNDSSIQGNCFIYNVKISGENFPPNGPVMQKKTQGWEPSTERLFARDGMLIGNDYMALK
                     LEGGGHYLCEFKSTYKAKKPVRMPGRHEIDRKLDVTSHNRDYTSVEQCEIAIARHSLLG
                     "
     misc_feature    3829..4417
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4300..4311)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4420..4436)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             4942..5122
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5154..5378
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5424..5852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5866..6195
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6046..6181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6243..6290
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6309..6683
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6813..6934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6983..6999)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7007..7023
                     /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(7031..7061)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7076..7097
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7385..7973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8144..9004)
                     /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(9005..9109)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta ataccgccat gcatctgcag agggccctgc gtatgagtgc
     2641 aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg
     2701 acccactgga caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg
     2761 ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag
     2821 tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga
     2881 cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg
     2941 ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc
     3001 atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag
     3061 tcgtgtcgtg cctgagagcg cagtctagag ccaccatggt gagtgtgatc gctaaacaaa
     3121 tgacctacaa ggtttatatg tcaggcacgg tcaatggaca ctactttgag gtcgaaggcg
     3181 atggaaaagg aaagccttac gagggagagc agacagtaaa gctcactgtc accaagggtg
     3241 gacctctgcc atttgcttgg gatattttat caccacagct tcagtacgga agcataccat
     3301 tcaccaagta ccctgaagac atccctgatt atttcaagca gtcattccct gagggatata
     3361 catgggagag gagcatgaac tttgaagatg gtgcagtgtg tactgtcagc aatgattcca
     3421 gcatccaagg caactgtttc atctacaatg tcaaaatctc tggtgagaac tttcctccca
     3481 atggacctgt tatgcagaag aagacacagg gctgggaacc cagcactgag cgtctctttg
     3541 cacgagatgg aatgctgata ggaaacgatt atatggctct gaagttggaa ggaggtggtc
     3601 actatttgtg tgaatttaaa tctacttaca aggcaaagaa gcctgtgagg atgccagggc
     3661 gccacgagat tgaccgcaaa ctggatgtaa ccagtcacaa cagggattac acatctgttg
     3721 agcagtgtga aatagccatt gcacgccact ctttgctcgg tggatccgcg atcgcctgat
     3781 ggcaacgtgg cgcgcccctg cagggaattc gatatcaagc ttatcgataa tcaacctctg
     3841 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3901 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3961 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4021 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4081 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4141 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4201 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     4261 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     4321 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4381 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgagac
     4441 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
     4501 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
     4561 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     4621 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac
     4681 acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca
     4741 ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc
     4801 aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg
     4861 gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca catggcccga
     4921 gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     4981 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     5041 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     5101 tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     5161 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     5221 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     5281 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     5341 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     5401 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5461 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5521 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5581 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5641 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5701 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5761 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5821 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     5881 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5941 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     6001 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     6061 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6121 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6181 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     6241 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     6301 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
     6361 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
     6421 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
     6481 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
     6541 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
     6601 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
     6661 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
     6721 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     6781 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     6841 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6901 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     6961 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7021 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7081 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7141 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7201 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7261 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7321 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7381 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7441 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7501 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7561 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7621 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7681 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7741 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7801 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     7861 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7921 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7981 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8041 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8101 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8161 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8221 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8281 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8341 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8401 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8461 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8521 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8581 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8641 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8701 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8761 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8821 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8881 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8941 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9001 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9061 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9121 gaaaagtgcc acctgac
//
