LOCUS       Exported                9391 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 9391)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Aug 29, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9391
                     /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)"
     CDS             3090..3998
                     /codon_start=1
                     /label=hCHIP
                     /translation="MKGKEEKEGGARLGAGGGSPEKSPSAQELKEQGNRLFVGRKYPEA
                     AACYGRAITRNPLVAVYYTNRALCYLKMQQHEQALADCRRALELDGQSVKAHFFLGQCQ
                     LEMESYDEAIANLQRAYSLAKEQRLNFGDDIPSALRIAKKKRWNSIEERRIHQESELHS
                     YLSRLIAAERERELEECQRNHEGDEDDSHVRAQQACIEAKHDKYMADMDELFSQVDEKR
                     KKRDIPDYLCGKISFELMREPCITPSGITYDRKDIEEHLQRVGHFDPVTRSPLTQEQLI
                     PNLAMKEVIDAFISENGWVEDY"
     CDS             4026..4055
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    4083..4671
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4554..4565)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4674..4690)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             5196..5376
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5408..5632
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5678..6106
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6120..6449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6300..6435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6497..6544
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6563..6937
                     /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    7067..7188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7237..7253)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7261..7277
                     /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(7285..7315)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7330..7351
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7639..8227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8398..9258)
                     /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(9259..9363)
                     /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 cagtctagaa tgaagggcaa ggaggagaag gagggcggcg
     3121 cacggctggg cgctggcggc ggaagccccg agaagagccc gagcgcgcag gagctcaagg
     3181 agcagggcaa tcgtctgttc gtgggccgaa agtacccgga ggcggcggcc tgctacggcc
     3241 gcgcgatcac ccggaacccg ctggtggccg tgtattacac caaccgggcc ttgtgctacc
     3301 tgaagatgca gcagcacgag caggccctgg ccgactgccg gcgcgccctg gagctggacg
     3361 ggcagtctgt gaaggcgcac ttcttcctgg ggcagtgcca gctggagatg gagagctatg
     3421 atgaggccat cgccaatctg cagcgagctt acagcctggc caaggagcag cggctgaact
     3481 tcggggacga catccccagc gctcttcgaa tcgcgaagaa gaagcgctgg aacagcattg
     3541 aggagcggcg catccaccag gagagcgagc tgcactccta cctctccagg ctcattgccg
     3601 cggagcgtga gagggagctg gaagagtgcc agcgaaacca cgagggtgat gaggacgaca
     3661 gccacgtccg ggcccagcag gcctgcattg aggccaagca cgacaagtac atggcggaca
     3721 tggacgagct tttttctcag gtggatgaga agaggaagaa gcgagacatc cccgactacc
     3781 tgtgtggcaa gatcagcttt gagctgatgc gggagccgtg catcacgccc agtggcatca
     3841 cctacgaccg caaggacatc gaggagcacc tgcagcgtgt gggtcatttt gaccccgtga
     3901 cccggagccc cctgacccag gaacagctca tccccaactt ggctatgaag gaggttattg
     3961 acgcattcat ctctgagaat ggctgggtgg aggactacag cggaccgacg cgtacgcggc
     4021 cgctcgagca gaaactcatc tcagaagagg atctgtgaga attcgatatc aagcttatcg
     4081 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4141 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4201 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     4261 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     4321 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     4381 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     4441 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     4501 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     4561 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     4621 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     4681 gtcgacctcg agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca
     4741 atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac
     4801 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     4861 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc
     4921 tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca ccagggccag
     4981 ggatcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagcaagaga
     5041 aggtagaaga agccaatgaa ggagagaaca cccgcttgtt acaccctgtg agcctgcatg
     5101 ggatggatga cccggagaga gaagtattag agtggaggtt tgacagccgc ctagcatttc
     5161 atcacatggc ccgagagctg catccggact gtactgggtc tctctggtta gaccagatct
     5221 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5281 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5341 tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac ccgctgatca
     5401 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     5461 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     5521 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     5581 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     5641 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     5701 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5761 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5821 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5881 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5941 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     6001 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     6061 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     6121 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6181 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6241 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     6301 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6361 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     6421 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     6481 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     6541 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     6601 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     6661 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     6721 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     6781 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     6841 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     6901 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
     6961 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     7021 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     7081 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     7141 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     7201 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7261 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7321 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7381 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7441 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7501 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7561 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7621 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7681 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7741 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7801 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7861 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7921 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7981 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8041 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     8101 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8161 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8221 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8281 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8341 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8401 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8461 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8521 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8581 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8641 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8701 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8761 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8821 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8881 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8941 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9001 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9061 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9121 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9181 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9241 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9301 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9361 gcgcacattt ccccgaaaag tgccacctga c
//
