LOCUS       pcDNA4.1-Myc-RNF        6579 bp DNA     circular SYN 22-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6579)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 22, 2024 from SnapGene 7.2.0
            https://www.snapgene.com
COMMENT     Sequence Label: pcDNA4.1-Myc-RNF8
FEATURES             Location/Qualifiers
     source          1..6579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2501..2530
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2546..2563
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2592..2816
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2862..3290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3304..3634
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3485..3620
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3682..3729
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3748..4122
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4252..4373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4422..4438)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4446..4462
                     /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(join(4470..4476,4477..4494,4495..4500))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4515..4536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4824..5412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5583..6374,6375..6443))
                     /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"
     promoter        complement(6444..6548)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggaacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attcatgggg gagcccggct tcttcgtcac aggagaccgc gccggtggcc ggagctggtg
     1081 cctgcggcgg gtggggatga gcgccgggtg gctgctgctg gaagatgggt gcgaggtgac
     1141 tgtaggacga ggatttggtg tcacatacca actggtatca aaaatctgcc ccctgatgat
     1201 ttctcgaaac cactgtgttt tgaagcagaa tcctgagggc caatggacaa ttatggacaa
     1261 caagagtcta aatggtgttt ggctgaacag agcgcgtctg gaacctttaa gggtctattc
     1321 cattcatcag ggagactaca tccaacttgg agtgcctctg gaaaataagg agaatgcgga
     1381 gtatgaatat gaagttactg aagaagactg ggagacaata tatccttgtc tttccccaaa
     1441 gaatgaccaa atgatagaaa aaaataagga attgagaact aaaaggaaat tcagtttgga
     1501 tgaattagca ggtcctggag ctgaaggccc ctcaaatttg aaatccaaaa taaataaagt
     1561 gtcttgtgaa tctggtcagc cagtgaaatc acaggggaaa ggtgaagtgg ccagtacacc
     1621 ctctgacaat ttggatccta agttgactgc ccttgagcca agtaagacca caggggctcc
     1681 catttaccct ggcttcccca aagtcacaga ggttcatcat gagcagaaag cctcaaactc
     1741 ttcagcatct cagagaagct tacagatgtt taaggtgacc atgtccagga ttctgaggct
     1801 caaaatacag atgcaggaaa aacatgaagc cgttatgaat gtgaaaaagc agacccaaaa
     1861 ggggaactca aagaaagttg tgcaaatgga gcaggaactt caggacttac agtcccagct
     1921 gtgtgcagag caggctcagc agcaggcaag agtggagcaa ctagagaaga ctttccagga
     1981 agaggaacag catcttcagg gtttggagat agcccaagga gaaaaggacc tgaagcaaca
     2041 gctggcccag gctctgcagg agcattgggc tctaatggaa gagctaaatc gcagcaagaa
     2101 ggactttgaa gcaatcattc aagccaagaa caaagaatta gagcagacca aggaagagaa
     2161 ggagaagatg caagcacaga aggaagaagt tcttagccac atgaatgatg tgctagagaa
     2221 tgagctccaa tgtattattt gttcagaata cttcattgag gctgtcacct tgaactgtgc
     2281 ccacagtttc tgctcctact gtatcaatga atggatgaag cggaagatag aatgccccat
     2341 ttgtcggaag gacattaagt ccaaaacgta ctctttggtt ctggacaatt gcattaataa
     2401 gatggtaaat aatctgagct cagaagtgaa agaacgacga attgttctca ttagggaacg
     2461 aaaagcaaag agattgttcc tcgagtctag agggcccttc gaacaaaaac tcatctcaga
     2521 agaggatctg aatatgcata ccggtcatca tcaccatcac cattgagttt aaacccgctg
     2581 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2641 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2701 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2761 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2821 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2881 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2941 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3001 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     3061 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3121 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3181 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     3241 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3301 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca
     3361 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3421 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3481 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3541 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3601 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3661 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     3721 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
     3781 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     3841 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     3901 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     3961 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     4021 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     4081 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
     4141 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4201 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4261 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4321 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4381 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4441 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4501 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4561 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4621 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4681 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4741 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4801 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4861 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4921 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4981 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     5041 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5101 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5161 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5221 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5281 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5341 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5401 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5461 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5521 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5581 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5641 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5701 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5761 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5821 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5881 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5941 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6001 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6061 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6121 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6181 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6241 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6301 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6361 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6421 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6481 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6541 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
