LOCUS       Exported                9232 bp ds-DNA     circular SYN 15-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9232)
  AUTHORS   Nike
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Sep 21, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9232
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(28..46)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(67..83)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    91..107
                     /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(115..145)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    160..181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(469..1057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1228..2088)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR-alt
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2089..2193)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2219..2674)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2816..2832
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2842..2860
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2881..3016
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    complement(3056..3177)
                     /label=SV40 poly(A) early  signal
                     /note="SV40 polyadenylation signal"
     LTR             3255..3488
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3560..4148
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4173..4865)
                     /codon_start=1
                     /label=mKate2Alt
                     /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHR"
     CDS             complement(4866..4919)
                     /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             complement(4920..5516)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR-alt
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(5529..6707)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     misc_feature    6846..6963
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7019..7267
                     /label=U6
     misc_feature    7366..7494
                     /label=gRNAscaffold
     misc_feature    7927..8160
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    8653..8778
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    complement(8825..9005)
                     /label=5LTR-zhike(TypeIISmutation)
     promoter        complement(9006..9232)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 ttaagcttgc agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
       61 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
      121 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
      181 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
      241 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctcat ccgcttcctc
      301 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
      361 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
      421 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      481 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      541 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
      601 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
      661 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      721 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      781 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
      841 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      901 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      961 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1021 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1081 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1141 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1201 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1261 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     1321 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc
     1381 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     1441 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     1501 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     1561 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     1621 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     1681 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     1741 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     1801 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     1861 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1921 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1981 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     2041 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     2101 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     2161 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     2221 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2281 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2341 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2401 gtcaaagggc gaaaaaccgt ctatcagggc gatcgcccac tacgtgaacc atcaccctaa
     2461 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2521 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2581 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2641 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     2701 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     2761 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     2821 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg tacgtcccta
     2881 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     2941 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     3001 gagttagggg cgggatgggc ggagttaggg gcgggatagc tagagccaga catgataaga
     3061 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3121 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttcct
     3181 ctcactctct gatattcatt tctttgcaag ttataaatac tgaataataa gatgacatga
     3241 actactatac gtactgctag agattttcca cactgactaa aagggtctga gggatctcta
     3301 gttaccagag tcacacaaca gacgggcaca cactacttga agcactcaag gcaagcttta
     3361 ttgaggctta agcagtgggt tccctagtta gccagagagc tcccaggctc agatctggtc
     3421 taaccagaga gagccagtac aagcaaaaag cagatcttgt catcgttggg agtgaattag
     3481 cccttccagt cccccctttt cttttaaaaa gtggctaaga tctacagctg ccttgtaagt
     3541 cattggtctt aaagtcgacg cggggaggcg gcccaaaggg agatccgact cgtctgaggg
     3601 cgaaggcgaa cacgcggaag aggccgcaga gccggcagca ggccgcggga aggaaggtcc
     3661 gctggattga gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca
     3721 acacaggcga gcagccaagg aaaggacgat gatttccccg acaacaccac ggaattgtca
     3781 gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc aggcggcgtt gagttccgcc
     3841 gtggcaatag ggagggggaa agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg
     3901 ccccaaccag tgggggttgc gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca
     3961 acgggccaca actcctcata aagagacagc aaccaggatt tatacaagga ggagaaaatg
     4021 aaagccatac gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc
     4081 gtaaaaggag caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga
     4141 ggttgattgt cgacttaacg cgtggatcct tatctgtgcc ccagtttgct agggaggtcg
     4201 cagtatctgg ccacagccac ctcgtgctgc tcgacgtagg tttctttgtc ggcctccttg
     4261 attctttcca gccttctgtc cacatagtag acgccgggca tcttgaggtt cttagcgggt
     4321 ttcttggatc tgtatgtggt tttcaagttg cagatcaggt ggcccccgcc cacgagcttc
     4381 agggccatgt cggctctgcc ttccaggccg ccgtcagcgg ggtacagggt ttcggtggag
     4441 gcctcccagc cgagtgtttt cttctgcatc acagggccgt tggatgggaa gttcacccct
     4501 ctgatcttga cgttgtagat gaggcagccg tcctggaggc tggtgtcctg ggtagcggtc
     4561 agcacgcccc cgtcctcgta tgtggtgact ctctcccatg tgaagccctc ggggaaggac
     4621 tgcttaaaga agtcggggat gccctgggtg tggttgatga aggttttgct gccgtacatg
     4681 aagctggtag ccaggatgtc gaaggcgaag gggagagggc cgccctcgac cgccttgatt
     4741 ctcatggtct gggtgccctc gtagggcttg ccttcgccct cggatgtgca cttgaagtgg
     4801 tggttgttca cggtgccctc catgtacagc ttcatgtgca tgttctcctt aatcagctcg
     4861 ctcacggggc cgggattttc ctccacgtcc ccgcatgtta gcagacttcc cctgccctcg
     4921 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
     4981 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtttccagg
     5041 aaggcgggca ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc
     5101 agacccttgc cctggtggtc gggcgagact ccgacggtgg ccaggaacca cgcgggctcc
     5161 ttgggccggt gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg
     5221 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
     5281 ggcgtggtcc agaccgccac ggcggcgccg tcgtccgcga cccacacctt gccgatgtcg
     5341 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
     5401 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
     5461 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggtg
     5521 gccgtacgtc acgacacctg aaatggaaga aaaaaacttt gaaccactgt ctgaggcttg
     5581 agaatgaacc aagatccaaa ctcaaaaagg gcaaattcca aggagaatta catcaagtgc
     5641 caagctggcc taacttcagt ctccacccac tcagtgtggg gaaactccaa cgcataaaac
     5701 ccctcccccc aacctaaaga cgacgtactc caaaagctcg agaactaatc gaggtgcctg
     5761 gacggcgccc ggtactccgt ggagtcacat gaagcgacgg ctgaggacgg aaaggccctt
     5821 ttcctttgtg tgggtgactc acccgcccgc tctcccgagc gccgcgtcct ccattttgag
     5881 ctccctgcag cagggccggg aagcggccat ctttccgctc acgcaactgg tgccgaccgg
     5941 gccagccttg ccgcccaggg cggggcgata cacggcggcg cgaggccagg caccagagca
     6001 ggccggccag cttgagacta cccccgtccg attctcggtg gccgcgctcg caggccccgc
     6061 ctcgccgaac atgtgcgctg ggacgcacgg gccccgtcgc cgcccgcggc cccaaaaacc
     6121 gaaataccag tgtgcagatc ttggcccgca tttacaagac tatcttgcca gaaaaaaagc
     6181 gtcgcagcag gtcatcaaaa attttaaatg gctagagact tatcgaaagc agcgagacag
     6241 gcgcgaaggt gccaccagat tcgcacgcgg cggccccagc gcccaggcca ggcctcaact
     6301 caagcacgag gcgaaggggc tccttaagcg caaggcctcg aactctccca cccacttcca
     6361 acccgaagct cgggatcaag aatcacgtac tgcagccagg tggaagtaat tcaaggcacg
     6421 caagggccat aacccgtaaa gaggccaggc ccgcgggaac cacacacggc acttacctgt
     6481 gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg cgactactgc acttatatac
     6541 ggttctcccc caccctcggg aaaaaggcgg agccagtaca cgacatcact ttcccagttt
     6601 accccgcgcc accttctcta ggcaccggat caattgccga cccctccccc caacttctcg
     6661 gggactgtgg gccatgtgcg ctctgcccac tgacgggcac cggagccaat tcccactcct
     6721 ttcaagacct agaaggtcca ttagctgcaa agattcctct ctgtttaaaa ctttatccat
     6781 ctttgcaccc gggccccctc gagccggcgc caaagtggat ctctgctgtc cctgtaataa
     6841 acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt
     6901 gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta
     6961 aaattgtgga tgaatactgc catttgtctc aagatctagt tacgccaagc ttggtaccga
     7021 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     7081 aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     7141 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     7201 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
     7261 aaacaccgga gacggttgta aagatatctg agcacacaaa atatctagaa tccatgctaa
     7321 aatattgttc ctgttccatt aactgcgcta aaaataaacg tctctgtttg agagctatgc
     7381 tggagcccga acagcatagc aagttcaaat aaggctagtc cgttatcaac ttggcccgaa
     7441 aaaaaagaaa aaaagaaaaa aagaaaaacg ggccaagtgg caccgagtcg gtgctttttt
     7501 tcaccgtgga atgcgagaat tctgggtccc ctcggggttg ggaggtgggt ctgaaacgat
     7561 aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa aactattctt
     7621 aaacctacca agcctcctac tatcattatg aataatttta tataccacag ccaatttgtt
     7681 atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc ttgttcattc
     7741 ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta
     7801 atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag agatttatta
     7861 ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga gcaaccccaa
     7921 atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct tgcctggagc
     7981 tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat agccctcagc
     8041 aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac cgtcagcgtc
     8101 attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa caaagctcct
     8161 attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt gggtgctact
     8221 cctaatggtt caatttttac tactttatat ttatataatt cacttctcca attgtccctc
     8281 atatctcctc ctccaggtct gaagatcagc ggccgcttgc tgtgcggtgg tcttactttt
     8341 gttttgctct tgctctatct tgtctaaagc ttccttggtg tcttttatct ctatcctttg
     8401 atgcacacaa tagagggttg ctactgtatt atataatgat ctaagttctt ctgatcctgt
     8461 ctgaagggat ggttgtagct gtcccagtat ttgtctacag ccttctgatg tttctaacag
     8521 gccaggatta actgcgaatc gttctagctc cctgcttgcc catactatat gttttaattt
     8581 atattttttc tttccccctg gccttaaccg aattttttcc caacgcgatc taattctccc
     8641 ccgcttaata ctgacgctct cgcacccatc tctctccttc tagcctccgc tagtcaaaat
     8701 ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg
     8761 agtcctgcgt cgagagagct cctctggttt ccctttcgct ttcaagtccc tgttcgggcg
     8821 ccactgctag agattttcca cactgactaa aagggtctga gggatctcta gttaccagag
     8881 tcacacaaca gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta
     8941 agcagtgggt tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga
     9001 gagccgttta tgtatcgagc taggcactta aatacaatat ctctgcaatg cggcaattca
     9061 gtggttcgtc caatccatgt cagacccgtc tgttgccttc ctaataaggc acgatcgtac
     9121 caccttactt ccaccaatcg gcatgcacgg tgctttttct ctccttgtaa ggcatgttgc
     9181 taactcatcg ttaccatgtt gcaagactac aagagtattg cataagacta ca
//
