LOCUS       Exported                9140 bp ds-DNA     circular SYN 02-JAN-2023
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9140)
  AUTHORS   Marek
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jan 11, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9140
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /locus_tag="HIV-1 Psi"
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    846..1079
                     /locus_tag="RRE"
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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    1489..1606
                     /locus_tag="cPPT/CTS"
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        1745..2923
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     CDS             2936..3532
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             3536..3589
                     /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"
     CDS             3602..4300
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     misc_feature    4315..4903
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4786..4797)
                     /codon_start=1
                     /locus_tag="Factor Xa site"
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        5059..5299
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     LTR             5479..5659
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5737..5858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5898..6033
                     /locus_tag="SV40 ori"
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6054..6072)
                     /locus_tag="T7 promoter"
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6082..6098)
                     /locus_tag="M13 fwd"
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6240..6628
                     /direction=RIGHT
                     /locus_tag="f1 ori(1)"
                     /label=f1 ori
                     /label=f1 ori(1)
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      6629..6668
                     /direction=RIGHT
                     /locus_tag="f1 ori"
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             6826..7686
                     /codon_start=1
                     /gene="bla"
                     /locus_tag="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7857..8445
                     /direction=RIGHT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8733..8754
                     /locus_tag="CAP binding site"
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8769..8799
                     /locus_tag="lac promoter"
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8807..8823
                     /locus_tag="lac operator"
                     /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     8831..8847
                     /locus_tag="M13 rev"
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8868..8886
                     /locus_tag="T3 promoter"
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8914..9140
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaatct tgagacaaat ggcagtattc atccacaatt ttaaaagaaa
     1501 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     1561 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     1621 ggacagcaga gatccacttt ggcgccggct cgagggggcc cgggtgcaaa gatggataaa
     1681 gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg
     1741 aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     1801 ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga
     1861 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     1921 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     1981 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     2041 aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg
     2101 gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc
     2161 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
     2221 tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt
     2281 ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg
     2341 ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct
     2401 gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt
     2461 gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc
     2521 accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg
     2581 gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt
     2641 tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct
     2701 cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc
     2761 gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat
     2821 gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca
     2881 gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg ccaccatgac
     2941 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3001 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3061 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3121 caaggtgtgg gtcgcggacg acggcgccgc cgtggcggtc tggaccacgc cggagagcgt
     3181 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3241 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     3301 gtggttcctg gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     3361 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     3421 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     3481 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg ccggagaggg
     3541 cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccac cggtcgccac
     3601 cagcgagctg attaaggaga acatgcacat gaagctgtac atggagggca ccgtggacaa
     3661 ccatcacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat
     3721 gagaatcaag gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctactag
     3781 cttcctctac ggcagcaaga ccttcatcaa ccacacccag ggcatccccg acttcttcaa
     3841 gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt
     3901 gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat
     3961 cagaggggtg aacttcacat ccaacggccc tgtgatgcag aagaaaacac tcggctggga
     4021 ggccttcacc gaaacgctgt accccgctga cggcggcctg gaaggcagaa acgacatggc
     4081 cctgaagctc gtgggcggga gccatctgat cgcaaacatc aagaccacat atagatccaa
     4141 gaaacccgct aagaacctca agatgcctgg cgtctactat gtggactaca gactggaaag
     4201 aatcaaggag gccaacaacg agacctacgt cgagcagcac gaggtggcag tggccagata
     4261 ctgcgacctc cctagcaaac tggggcacaa gcttaattaa cgcgttaagt cgacaatcaa
     4321 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4381 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4441 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4501 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4561 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4621 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4681 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     4741 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4801 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4861 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa
     4921 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     4981 ggctaattca ctcccaacga agacaagatt tcaccattat cgtttcagac ccacctccca
     5041 accccgaggg gacccagaga gggcctattt cccatgattc cttcatattt gcatatacga
     5101 tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta
     5161 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     5221 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     5281 tatatcttgt ggaaaggacg aaacaccgga gacggttgtt aattaatgta cgtctctgtt
     5341 taagagctat gctggaaaca gcatagcaag tttaaataag gctagtccgt tatcaacttg
     5401 aaaaagtggc accgagtcgg tgcttttttt gaattcttgg cgtaactaga tcttgagaca
     5461 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5521 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5581 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5641 gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta
     5701 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     5761 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5821 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     5881 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5941 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     6001 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     6061 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     6121 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     6181 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     6241 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     6301 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     6361 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     6421 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     6481 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     6541 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     6601 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6661 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     6721 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6781 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6841 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6901 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6961 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     7021 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     7081 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     7141 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     7201 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     7261 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7321 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     7381 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     7441 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7501 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7561 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7621 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7681 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7741 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7801 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7861 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7921 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7981 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     8041 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     8101 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     8161 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     8221 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     8281 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     8341 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     8401 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8461 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     8521 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     8581 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     8641 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     8701 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     8761 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     8821 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     8881 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
     8941 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
     9001 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
     9061 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
     9121 gcctagctcg atacataaac
//
