LOCUS       Exported                9933 bp DNA     circular SYN 16-MAY-2025
DEFINITION  Exported.
ACCESSION   V013504
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9933)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9933)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    680..805
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1302..1535
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1720..1764
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
     CDS             1913..1954
                     /label=Protein Tat
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2026..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        2200..2579
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2581..2858
                     /label=chicken beta-actin promoter
     intron          2859..3876
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    3965..4551
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4553..5269
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             5288..5887
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    5901..6489
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6696..7329
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7457..7473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7481..7497
                     /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(7505..7535)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7550..7571)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7859..8447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8618..9478)
                     /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(9479..9583)
                     /label=AmpR promoter
     polyA_signal    9631..9765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             join(9933,1..633)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga tctaccacac
       61 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
      121 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
      181 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
      241 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
      301 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
      361 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
      421 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
      481 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
      541 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
      601 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
      661 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
      721 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
      781 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
      841 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
      901 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
      961 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     1021 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     1081 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     1141 agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     1201 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1261 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1321 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1381 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1441 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1501 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1561 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1621 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1681 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1741 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1801 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1861 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1921 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1981 agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccttta aaagaaaagg
     2041 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2101 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2161 cagcagagat ccagtttatc gataagcttg ggagttccgg acattgatta ttgactagtt
     2221 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     2281 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     2341 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     2401 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     2461 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     2521 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     2581 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
     2641 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
     2701 ggcccccccc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggaaaggtg
     2761 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
     2821 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgttgcg cgctgccttc
     2881 cccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
     2941 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
     3001 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
     3061 gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg
     3121 ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt
     3181 gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg
     3241 ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg
     3301 gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc
     3361 acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg
     3421 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
     3481 gctcggggga aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     3541 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     3601 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     3661 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     3721 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     3781 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     3841 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     3901 gtgctgtctc atcattttgg caaagaattc ctcgagacta gttctagagc ggccgcggat
     3961 cccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     4021 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     4081 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     4141 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     4201 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     4261 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     4321 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     4381 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     4441 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     4501 acggggacgt ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccatggtgag
     4561 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     4621 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     4681 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     4741 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     4801 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     4861 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     4921 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     4981 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     5041 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     5101 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     5161 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     5221 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaaga caaggagacg
     5281 accttccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg
     5341 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     5401 cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     5461 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     5521 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     5581 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     5641 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     5701 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     5761 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     5821 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     5881 tgcctagacg cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg
     5941 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     6001 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     6061 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     6121 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     6181 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     6241 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     6301 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     6361 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     6421 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     6481 cctccccgcc tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag
     6541 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
     6601 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
     6661 agatcttagc cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg
     6721 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca
     6781 gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct
     6841 agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt
     6901 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt
     6961 tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa
     7021 ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg
     7081 cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg
     7141 cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     7201 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     7261 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     7321 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     7381 gaatatcaga gagtgagagg ccttgacatt gctagcgttt accgtcgacc tctagctaga
     7441 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7501 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7561 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7621 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7681 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7741 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     8041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     8101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     8161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     8221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     8281 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     8341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     8401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     8461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     8521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     8581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     8641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     8701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     8761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     8941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     9001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     9061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     9121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     9181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     9241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     9301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     9361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     9421 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9481 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     9541 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     9601 tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta taatggttac
     9661 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     9721 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa
     9781 gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg
     9841 tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt
     9901 atttgttaaa tatgtactac aaacttagta gtt
//
