LOCUS       Exported                8786 bp ds-DNA     circular SYN 23-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8786)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jul 1, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8786
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    139..160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        175..205
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    213..229
                     /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)."
     primer_bind     237..253
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        274..292
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        320..546
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             547..727
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    774..899
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1392..1625
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1810..1854
                     /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    2121..2237
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2260..2563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2564..2767
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3858..3878
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3879..3944
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(3997..4021)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4052..4640
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4523..4534)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4696..5025
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4876..5011
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5073..5120
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5139..5537
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5624..5857
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5929..6050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6090..6225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6246..6264)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6274..6290)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6432..6887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6913..7017
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7018..7878
                     /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"
     rep_origin      8049..8637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
       61 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      121 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      181 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      241 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
      301 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta
      361 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
      421 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
      481 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
      541 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      601 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
      661 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
      721 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
      781 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
      841 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
      901 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
      961 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     1021 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     1081 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     1141 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     1201 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     1261 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     1321 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     1381 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     1441 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     1501 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     1561 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     1621 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     1681 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     1741 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     1801 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     1861 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     1921 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     1981 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     2041 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     2101 tctcgacggt atcggttaac ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     2161 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     2221 acaaaattca aaattttatc gataagcttg ggagttccgc gttacataac ttacggtaaa
     2281 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     2341 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     2401 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     2461 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     2521 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     2581 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     2641 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     2701 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     2761 cagagctcgt ttagtgaacc gtcagatcag tccaaggtac agtcgccgcg tgcggagctt
     2821 gttactggtt acttggcctc atggcggtcc gagcttcgtt cgagaacaac tgtgagatcg
     2881 gctgctttgc caagctcacc aacacctact gtctggtagc gatcggaggc tcagagaact
     2941 tctacagtgt gttcgagggc gagctctccg ataccatccc cgtggtgcac gcgtctatcg
     3001 ccggctgccg catcatcggg cgcatgtgtg tggggaacag gcacggtctc ctggtaccca
     3061 acaataccac cgaccaggag ctgcaacaca ttcgcaacag cctcccagac acagtgcaga
     3121 ttaggcgggt ggaggagcgg ctctcagcct tgggcaatgt caccacctgc aatgactacg
     3181 tggccttggt ccacccagac ttggacaggg agacagaaga aattctggca gatgtgctca
     3241 aggtggaagt cttcagacag acagtggccg accaggtgct agtaggaagc tactgtgtct
     3301 tcagcaatca gggagggctg gtgcatccca agacttcaat tgaagaccag gatgagctgt
     3361 cctctcttct tcaagtcccc cttgtggcgg ggactgtgaa ccgaggcagt gaggtgattg
     3421 ctgctgggat ggtggtgaat gactggtgtg ccttctgtgg cctggacaca accagcacag
     3481 agctgtcagt ggtggagagt gtcttcaagc tgaatgaagc ccagcctagc accattgcca
     3541 ccagcatgcg ggattccctc attgacagcc tcacctgagt caccttccaa gttgttccat
     3601 gggctcctgg ctctggactg tggccaacct tctccacatt ccgcccaatc tgtaccggat
     3661 gctggcaggg aggtggcaga gagctcactg ggactgaggg gctgggcacc caaccctttt
     3721 ccacctgtgc ttatcgcctg gatctatcat tactgcaaaa acctgctctg ttgtgctggc
     3781 tggcaggccc tgtggctgct ggctgagggt tctgctgtcc tgtgccaccc cattaaagtg
     3841 cagttccctc cgggccagag aacctgtact tccagggcga ctacaaagac catgacggtg
     3901 attataaaga tcatgacatc gattacaagg atgacgatga caagtagaaa cactaggatc
     3961 cggtaccgaa ttcgcggccg cactcgagat atctagaccc agctttcttg tacaaagtgg
     4021 ttgatatcca gcacagtggc ggccgctcga caatcaacct ctggattaca aaatttgtga
     4081 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     4141 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     4201 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     4261 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     4321 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     4381 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     4441 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     4501 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     4561 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     4621 cctttgggcc gcctccccgc ctggaattct gcagatatcc ggttagtaat gagtttggaa
     4681 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4741 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4801 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4861 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4921 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4981 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     5041 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
     5101 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagcct ttgtctcaag
     5161 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
     5221 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
     5281 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
     5341 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     5401 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
     5461 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     5521 atgtgtggga gggctaagca caattcgagc tcggtacctt taagaccaat gacttacaag
     5581 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctagctcac
     5641 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     5701 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5761 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5821 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     5881 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     5941 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6001 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6061 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6121 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6181 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
     6241 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     6301 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     6361 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     6421 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6481 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6541 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6601 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6661 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6721 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6781 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6841 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     6901 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6961 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     7021 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     7081 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     7141 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     7201 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     7261 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     7321 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     7381 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     7441 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     7501 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     7561 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7621 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7681 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7741 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7801 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7861 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7921 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7981 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     8041 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     8101 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     8161 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     8221 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     8281 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     8341 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     8401 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     8461 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     8521 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     8581 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     8641 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     8701 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     8761 accgctcgcc gcagccgaac gaccga
//
