LOCUS       Exported                8347 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 8347)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jan 11, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     >Mutate T9886A and A9907T
FEATURES             Location/Qualifiers
     source          1..8347
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    78..199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      239..374
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(395..413)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(423..439)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      581..1036
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1062..1166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1167..2027
                     /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      2122..2804
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
     rep_origin      2198..2786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3074..3095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3110..3140
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_binding    3144..3166
                     /locus_tag="LacO"
                     /label=LacO
     protein_bind    3148..3164
                     /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     3172..3188
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3209..3227
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3255..3481
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3482..3662
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3709..3834
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4327..4560
                     /locus_tag="RRE"
                     /label=RRE
     CDS             4745..4789
                     /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"
     promoter        4967..5207
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    5393..5510
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5649..6826
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     CDS             6839..7438
                     /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    7454..8042
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             8114..8347
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
ORIGIN
        1 gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
       61 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
      121 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
      181 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
      241 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
      301 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
      361 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
      421 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
      481 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
      541 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
      601 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
      661 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
      721 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
      781 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
      841 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
      901 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      961 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1021 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1081 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1141 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1201 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1261 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1321 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1381 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1441 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1501 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1561 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1621 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1681 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1741 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1801 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1861 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1921 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1981 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2041 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2101 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2161 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2221 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2281 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2341 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2401 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2461 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2521 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2581 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2641 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2701 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2761 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2821 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2881 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2941 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     3001 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     3061 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     3121 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     3181 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     3241 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     3301 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     3361 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     3421 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
     3481 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     3541 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     3601 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     3661 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     3721 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     3781 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     3841 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     3901 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     3961 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     4021 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     4081 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     4141 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     4201 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     4261 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     4321 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     4381 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     4441 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     4501 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     4561 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     4621 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     4681 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     4741 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     4801 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     4861 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     4921 accattatcg tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc
     4981 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa
     5041 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
     5101 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
     5161 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaga
     5221 cggttgttaa ttaatgtacg tctctgttta agagctatgc tggaaacagc atagcaagtt
     5281 taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa
     5341 ttcaagcttg gcgtaactag atcttgagac aaatggcagt attcatccac aattttaaaa
     5401 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     5461 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     5521 acagggacag cagagatcca ctttggcgcc ggctcgaggg ggcccgggtg caaagatgga
     5581 taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag
     5641 tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     5701 agttgggggg aggggtcggc aattgatccg gtgcctagag aaggtggcgc ggggtaaact
     5761 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     5821 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
     5881 taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc
     5941 cttgaattac ttccactggc tgcagtacgt gattcttgat cccgagcttc gggttggaag
     6001 tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga
     6061 ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct
     6121 cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt
     6181 tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt
     6241 tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg
     6301 cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct
     6361 ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc
     6421 ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa
     6481 atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc
     6541 ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca
     6601 cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta
     6661 tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt
     6721 gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc
     6781 tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgacgta cggccaccat
     6841 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     6901 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     6961 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     7021 cggcaaggtg tgggtcgcgg acgacggcgc cgccgtggcg gtctggacca cgccggagag
     7081 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     7141 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     7201 cgcgtggttc ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag
     7261 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     7321 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     7381 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaac
     7441 gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     7501 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     7561 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     7621 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     7681 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     7741 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     7801 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     7861 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     7921 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     7981 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     8041 gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     8101 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     8161 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     8221 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     8281 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     8341 tctagca
//
