LOCUS       pLJM1-Halo-NLS          8302 bp DNA     circular SYN 11-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8302)
  AUTHORS   Azra Lari
  TITLE     Direct Submission
  JOURNAL   Exported Dec 11, 2025 from SnapGene 6.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          399..1283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        131..334
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      390..395
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             396..1283
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEIS"
     regulatory      396..398
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1284..1319
                     /codon_start=1
                     /label=GSA linker
                     /translation="GSAGSAAGSGEF"
     CDS             1320..1340
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1385..1502
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1551..2061
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             2083..2682
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             2810..3043
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3115..3236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3276..3411
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3432..3450)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3460..3476)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3618..4073
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4099..4203
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4204..5064
                     /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      5235..5823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6111..6132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6147..6177
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6185..6201
                     /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     6209..6225
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6246..6264
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6292..6518
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6519..6699
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6746..6871
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7364..7597
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7782..7826
                     /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"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcgctaccgg tgtcgacgat atctctagag ccaccatggc agaaatcggt actggctttc
      421 cattcgaccc ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc
      481 cgcgcgatgg cacccctgtg ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc
      541 gcaacatcat cccgcatgtt gcaccgaccc atcgctgcat tgctccagac ctgatcggta
      601 tgggcaaatc cgacaaacca gacctgggtt atttcttcga cgaccacgtc cgcttcatgg
      661 atgccttcat cgaagccctg ggtctggaag aggtcgtcct ggtcattcac gactggggct
      721 ccgctctggg tttccactgg gccaagcgca atccagagcg cgtcaaaggt attgcattta
      781 tggagttcat ccgccctatc ccgacctggg acgaatggcc agaatttgcc cgcgagacct
      841 tccaggcctt ccgcaccacc gacgtcggcc gcaagctgat catcgatcag aacgttttta
      901 tcgagggtac gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt
      961 accgcgagcc gttcctgaat cctgttgacc gcgagccact gtggcgcttc ccaaacgagc
     1021 tgccaatcgc cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac atggactggc
     1081 tgcaccagtc ccctgtcccg aagctgctgt tctggggcac cccaggcgtt ctgatcccac
     1141 cggccgaagc cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg gacatcggcc
     1201 cgggtctgaa tctgctgcaa gaagacaacc cggacctgat cggcagcgag atcgcgcgct
     1261 ggctgtcgac gctcgagatt tccggatccg ctggctccgc tgctggttct ggcgaattcc
     1321 caaagaagaa gcggaaggtc tgagaattct cgacctcgag acaaatggca gtattcatcc
     1381 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     1441 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     1501 ttcgggttta ttacagggac agcagagatc cactttggcc gcggctcgag ggggttgggg
     1561 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
     1621 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
     1681 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
     1741 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
     1801 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
     1861 cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa
     1921 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
     1981 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
     2041 atcaccgacc tctctcccca gggggatcca ccggagctta ccatgaccga gtacaagccc
     2101 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     2161 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     2221 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     2281 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     2341 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     2401 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     2461 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     2521 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     2581 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     2641 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg
     2701 cagcgcccga ccgaaaggag cgcacgaccc catgcatcgg tacctttaag accaatgact
     2761 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     2821 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     2881 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     2941 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     3001 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     3061 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     3121 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3181 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3241 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3301 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3361 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     3421 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     3481 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3541 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3601 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3661 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3721 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3781 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3841 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3901 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3961 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4021 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     4081 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4141 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4201 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4261 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4321 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4381 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4441 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4501 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4561 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4621 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4681 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4741 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4801 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4861 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4921 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4981 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5041 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5101 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5161 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5221 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5281 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5341 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5401 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5461 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5521 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5581 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5641 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5701 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5761 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5821 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5881 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5941 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6001 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6061 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     6121 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     6181 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     6241 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
     6301 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     6361 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
     6421 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     6481 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     6541 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6601 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     6661 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     6721 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     6781 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     6841 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     6901 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     6961 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     7021 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     7081 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     7141 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     7201 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     7261 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     7321 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     7381 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     7441 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     7501 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     7561 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     7621 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     7681 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     7741 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     7801 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     7861 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     7921 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     7981 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     8041 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgatca cgagactagc
     8101 ctcgagcggc cgcccccttc accgagggcc tatttcccat gattccttca tatttgcata
     8161 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
     8221 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     8281 tatgttttaa aatggactat ca
//
