LOCUS       Exported                8630 bp ds-DNA     circular SYN 15-DEC-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8630)
  AUTHORS   jay
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Apr 24, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8630
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             236..413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1835..1952
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2017..3228
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      3261..3270
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3267..3983
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3993..4049
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4062..4358
                     /codon_start=1
                     /label=NRL-205 (TR)
                     /translation="MSVRELNRQLRGCGRDEALRLKQRRRTLKNRGYAQACRSKRLQQR
                     RGLEAERARLAAQLDALRAEVARLARERDLYKARCDRLTSSGPGSGDPSHLFL"
     CDS             4374..4427
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    4471..5059
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4942..4953)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5150..5383
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5455..5576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5616..5751
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5772..5790)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5800..5816)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5961..6416
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6445..6549
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6550..7410
                     /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      7581..8169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8457..8478
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8493..8523
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8531..8547
                     /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     8555..8571
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8592..8610
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacaa taaacgggtc
      241 tctctggtta gaccagatct gagcctggct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acctgaaagc gaaagggaaa ccagagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc
     1801 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     1861 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     1921 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     1981 cagtttggtt aattaacccg tgtcggctcc agatctggcc tccgcgccgg gttttggcgc
     2041 ctcccgcggg cgcccccctc ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg
     2101 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     2161 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     2221 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     2281 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     2341 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     2401 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     2461 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     2521 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     2581 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     2641 atggtgggcg gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa agctcttatt
     2701 cgggtgagat gggctggggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     2761 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     2821 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     2881 tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg
     2941 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     3001 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     3061 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     3121 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     3181 agtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacga agcttgggct
     3241 gcaggtccga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     3301 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3361 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3421 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     3481 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3541 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3601 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3661 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3721 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     3781 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3841 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     3901 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3961 tcggcatgga cgagctgtac aagggatctg gagcaacaaa cttctcacta ctcaaacaag
     4021 caggtgacgt ggaggagaat cccgggcctt ctagaggatc catgtctgtg cgggagctaa
     4081 accggcagct gcggggctgc gggcgcgacg aggcgctgcg gctgaagcag aggcgccgca
     4141 cgctgaagaa ccgcggctac gcgcaggcct gtcgctccaa gcggctgcag cagcggcgcg
     4201 ggctggaggc cgagcgcgcc cgcctggccg cccagctgga cgcgctgcgg gccgaggtgg
     4261 cccgcctggc ccgggagcgc gatctctaca aggctcgctg tgaccggcta acctcgagcg
     4321 gccccgggtc cggggacccc tcccacctct tcctctgagg atccggctcc ggcgagggca
     4381 ggggaagtct actaacatgc ggggacgtgg aggaaaatcc cggcccagct agcgtgtgat
     4441 cagaattcgt tccggagtcg tcgactcgac aatcaacctc tggattacaa aatttgtgaa
     4501 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     4561 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     4621 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     4681 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     4741 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4801 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4861 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4921 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4981 ctgccggctc tgcggcctct tccgcatctt cgccttcgcc ctcagacgag tcggatctcc
     5041 ctttgggccg cctccccgcc tggaattaat tcgagctcgg tacctttaag accaatgact
     5101 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     5161 cgtaactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     5221 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     5281 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     5341 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     5401 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     5461 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5521 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5581 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5641 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5701 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     5761 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     5821 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     5881 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     5941 cctgaatggc gaatggcgcg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6001 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6061 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6121 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6181 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6241 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     6301 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6361 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caatttccca
     6421 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6481 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6541 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6601 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6661 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6721 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6781 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6841 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6901 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6961 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7021 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7081 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7141 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7201 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7261 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7321 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7381 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7441 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7501 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7561 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7621 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7681 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     7741 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7801 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7861 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7921 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7981 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8041 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8101 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8161 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8221 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     8281 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     8341 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     8401 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     8461 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     8521 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     8581 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
//
