LOCUS       Exported                9314 bp ds-DNA     circular SYN 26-FEB-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 9314)
  AUTHORS   Silvie Trantirkova
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Feb 26, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9314
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1296..1320
                     /label=SEQ_CMV_FW
     enhancer        1360..1724
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1725..1928
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    1930..1948
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1951..1969
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    2010..2036
                     /label=SEQ_FW_PBtetr
     CDS             2091..2552
                     /codon_start=1
                     /label=hSOD
                     /translation="MATKAVCVLKGDGPVQGIINFEQKESNGPVKVWGSIKGLTEGLHG
                     FHVHEFGDNTAGCTSAGPHFNPLSRKHGGPKDEERHVGDLGNVTADKDGVADVSIEDSV
                     ISLSGDHCIIGRTLVVHEKADDLGKGGNEESTKTGNAGSRLACGVIGIAQ"
     CDS             2553..2576
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2589..3175
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3188..3946
                     /label=copGFP
     promoter        3995..4206
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             4219..4487
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     misc_feature    complement(4446..4470)
                     /label=SEQ_TETr_FW
     CDS             4524..5141
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG"
     CDS             5178..5231
                     /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             5238..5837
                     /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    5281..5305
                     /label=SEQ_TETr_RV
     polyA_signal    5884..6005
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7293..7309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7317..7333
                     /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(7341..7371)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7386..7407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7695..8283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8454..9314)
                     /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"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccagac tagttattaa tagtaatcaa
     1381 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     1441 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     1501 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     1561 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     1621 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     1681 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     1741 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     1801 ttgacgtcaa tgggagtttg ttttggaacc aaaatcaacg ggactttcca aaatgtcgta
     1861 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     1921 gcagagctct ccctatcagt gatagagatc tccctatcag tgatagagat cgtcgacgag
     1981 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     2041 gaagacaccg ggaccgatcc agcctccgga ctctagcggt ctaggctagc atggcgacga
     2101 aggccgtgtg cgtgctgaag ggcgacggcc cagtgcaggg catcatcaat ttcgagcaga
     2161 aggaaagtaa tggaccagtg aaggtgtggg gaagcattaa aggactgact gaaggcctgc
     2221 atggattcca tgttcatgag tttggagata atacagcagg ctgtaccagt gcaggtcctc
     2281 actttaatcc tctatccaga aaacacggtg ggccaaagga tgaagagagg catgttggag
     2341 acttgggcaa tgtgactgct gacaaagatg gtgtggccga tgtgtctatt gaagattctg
     2401 tgatctcact ctcaggagac cattgcatca ttggccgcac actggtggtc catgaaaaag
     2461 cagatgactt gggcaaaggt ggaaatgaag aaagtacaaa gacaggaaac gctggaagtc
     2521 gtttggcttg tggtgtaatt gggatcgccc aagactacaa ggacgacgac gacaagtgag
     2581 cggccgcgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     2641 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     2701 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     2761 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     2821 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     2881 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     2941 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     3001 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     3061 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     3121 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggc
     3181 gtccggaatg gagagcgacg agagcggcct gcccgccatg gagatcgagt gccgcatcac
     3241 cggcaccctg aacggcgtgg agttcgagct ggtgggcggc ggagagggca cccccaagca
     3301 gggccgcatg accaacaaga tgaagagcac caaaggcgcc ctgaccttca gcccctacct
     3361 gctgagccac gtgatgggct acggcttcta ccacttcggc acctacccca gcggctacga
     3421 gaaccccttc ctgcacgcca tcaacaacgg cggctacacc aacacccgca tcgagaagta
     3481 cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc tacgaggccg gccgcgtgat
     3541 cggcgacttc aaggtggtgg gcaccggctt ccccgaggac agcgtgatct tcaccgacaa
     3601 gatcatccgc agcaacgcca ccgtggagca cctgcacccc atgggcgata acgtgctggt
     3661 gggcagcttc gcccgcacct tcagcctgcg cgacggcggc tactacagct tcgtggtgga
     3721 cagccacatg cacttcaaga gcgccatcca ccccagcatc ctgcagaacg ggggccccat
     3781 gttcgccttc cgccgcgtgg aggagctgca cagcaacacc gagctgggca tcgtggagta
     3841 ccagcacgcc ttcaagaccc ccatcgcctt cgccagatcc cgcgctcagt cgtccaattc
     3901 tgccgtggac ggcaccgccg gacccggctc caccggatct cgctagagct gaatctaagt
     3961 cgacggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     4021 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     4081 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     4141 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     4201 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     4261 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     4321 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     4381 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     4441 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     4501 gcgcctactc tagagccgcc accatgtcca gattagataa aagtaaagtg attaacagcg
     4561 cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga
     4621 agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg
     4681 acgccttagc cattgagatg ttagataggc accatactca cttttgccct ttagaagggg
     4741 aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc
     4801 atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc
     4861 tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg
     4921 cactcagcgc tgtggggcat tttactttag gttgcgtatt ggaagatcaa gagcatcaag
     4981 tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag
     5041 ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga
     5101 tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtccgcgtac agcggatccc
     5161 gggagttcag atcttatgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga
     5221 atcccggccc tgaattcatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     5281 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     5341 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     5401 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     5461 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     5521 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     5581 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     5641 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     5701 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     5761 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     5821 gcaagcccgg tgcctgagtc gacaatcaac ctctggatta caaaatttgt ggttaactaa
     5881 ctaaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5941 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6001 tcttatcatg tctggaattg actcaaatga tgtcaattag tctatcagaa gctcatctgg
     6061 tctcccttcc gggggacaag acatccctgt ttaatattta aacagcagtg ttcccaaact
     6121 gggttcttat atcccttgct ctggtcaacc aggttgcagg gtttcctgtc ctcacaggaa
     6181 cgaagtccct aaagaaacag tggcagccag gtttagcccc ggaattgact ggattccttt
     6241 tttagggccc attggtatgg ctttttcccc gtatcccccc aggtgtctgc aggctcaaag
     6301 agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt
     6361 ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg
     6421 cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg
     6481 gggggctgtc cctgatatct ataacaagaa aatatatata taataagtta tcacgtaagt
     6541 agaacatgaa ataacaatat aattatcgta tgagttaaat cttaaaagtc acgtaaaaga
     6601 taatcatgcg tcattttgac tcacgcggtc gttatagttc aaaatcagtg acacttaccg
     6661 cattgacaag cacgcctcac gggagctcca agcggcgact gagatgtcct aaatgcacag
     6721 cgacggattc gcgctattta gaaagagaga gcaatatttc aagaatgcat gcgtcaattt
     6781 tacgcagact atctttctag ggttaatcta gctgcatcag gatcatatcg tcgggtcttt
     6841 tttccggctc agtcatcgcc caagctggcg ctatctgggc atcggggagg aagaagcccg
     6901 tgccttttcc cgcgaggttg aagcggcatg gaaagagttt gccgaggatg actgctgctg
     6961 cattgacgtt gagcgaaaac gcacgtttac catgatgatt cgggaaggtg tggccatgca
     7021 cgcctttaac ggtgaactgt tcgttcaggc cacctgggat accagttcgt cgcggctttt
     7081 ccggacacag ttccggatgg tcagcccgaa gcgcatcagc aacccgaaca ataccggcga
     7141 cagccggaac tgccgtgccg gtgtgcagat taatgacagc ggtgcggcgc tgggatatta
     7201 cgtcagcgag gacgggtatc ctggctggat gccgcagaaa tggacatgga taccccgtga
     7261 gttacccggc gggcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     7321 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     7381 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     7441 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7501 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7561 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7621 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7681 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7741 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7801 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7861 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7921 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7981 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     8041 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     8101 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     8161 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     8221 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     8281 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     8341 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     8401 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     8461 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     8521 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8581 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8641 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8701 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8761 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8821 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8881 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8941 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     9001 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     9061 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     9121 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     9181 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     9241 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     9301 tgttgaatac tcat
//
