LOCUS       pEPI-1.        6944 bp DNA     circular SYN 14-JUN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pEPI-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  AUTHORS   caoheibi
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             553..1269
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    1276..3476
                     /label=S/MAR
                     /note="S/MAR"
                     /note="S/MARs-element(Scaffold or Matrix Attachment
                     Regions) are typically 70% A/T-rich sequences and are often
                     found in association with chromosomal origins of 
                     bidirectional replication. It could be used to construct 
                     non-viral episomal vectors in mammalian cells."
     misc_feature    3483..3548
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3672..3793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3800..4255)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4282..4386
                     /label=AmpR promoter
     promoter        4388..4745
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4780..5571
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5806..5853
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6182..6770
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6885..6944
                     /label=CMV
                     /note="Human cytomegalovirus immediate early
                     enhancer/promoter"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagg aattcgaatt cagcaaggtc gccacgcaca agatcaatat taacaatcag
     1321 tcatctctct ttagcaataa aaaggtgaaa aattacattt taaaaatgac accatagacg
     1381 atgtatgaaa ataatctact tggaaataaa tctaggcaaa gaagtgcaag actgttaccc
     1441 agaaaactta caaattgtaa atgagaggtt agtgaagatt taaatgaatg aagatctaaa
     1501 taaacttata aattgtgaga gaaattaatg aatgtctaag ttaatgcaga aacggagaga
     1561 catactatat tcatgaacta aaagacttaa tattgtgaag gtatactttc ttttcacata
     1621 aatttgtagt caatatgttc accccaaaaa agctgtttgt taacttgtca acctcatttc
     1681 aaaatgtata tagaaagccc aaagacaata acaaaaatat tcttgtagaa caaaatggga
     1741 aagaatgttc cactaaatat caagatttag agcaaagcat gagatgtgtg gggatagaca
     1801 gtgaggctga taaaatagag tagagctcag aaacagaccc attgatatat gtaagtgacc
     1861 tatgaaaaaa atatggcatt ttacaatggg aaaatgatga tctttttctt ttttagaaaa
     1921 acagggaaat atatttatat gtaaaaaata aaagggaacc catatgtcat accatacaca
     1981 caaaaaaatt ccagtgaatt ataagtctaa atggagaagg caaaacttta aatcttttag
     2041 aaaataatat agaagcatgc catcatgact tcagtgtaga gaaaaatttc ttatgactca
     2101 aagtcctaac cacaaagaaa agattgttaa ttagattgca tgaatattaa gacttatttt
     2161 taaaattaaa aaaccattaa gaaaagtcag gccatagaat gacagaaaat atttgcaaca
     2221 ccccagtaaa gagaattgta atatgcagat tataaaaaga agtcttacaa atcagtaaaa
     2281 aataaaacta gacaaaaatt tgaacagatg aaagagaaac tctaaataat cattacacat
     2341 gagaaactca atctcagaaa tcagagaact atcattgcat atacactaaa ttagagaaat
     2401 attaaaaggc taagtaacat ctgtggcaat attgatggta tataaccttg atatgatgtg
     2461 atgagaacag tactttaccc catgggcttc ctccccaaac ccttacccca gtataaatca
     2521 tgacaaatat actttaaaaa ccattaccct atatctaacc agtactcctc aaaactgtca
     2581 aggtcatcaa aaataagaaa agtctgagga actgtcaaaa ctaagaggaa cccaaggaga
     2641 catgagaatt atatgtaatg tggcattctg aatgagatcc cagaacagaa aaagaacagt
     2701 agctaaaaaa ctaatgaaat ataaataaag tttgaacttt agtttttttt aaaaaagagt
     2761 agcattaaca cggcaaagtc attttcatat ttttcttgaa cattaagtac aagtctataa
     2821 ttaaaaattt tttaaatgta gtctggaaca ttgccagaaa cagaagtaca gcagctatct
     2881 gtgctgtcgc ctaactatcc atagctgatt ggtctaaaat gagatacatc aacgctcctc
     2941 catgtttttt gttttctttt taaatgaaaa actttatttt ttaagaggag tttcaggttc
     3001 atagcaaaat tgagaggaag gtacattcaa gctgaggaag ttttcctcta ttcctagttt
     3061 actgagagat tgcatcatga atgggtgtta aattttgtca aatgcttttt ctgtgtctat
     3121 caatatgacc atgtgatttt cttctttaac ctgttgatgg gacaaattac gttaattgat
     3181 tttcaaacgt tgaaccaccc ttacatatct ggaataaatt ctacttggtt gtggtgtata
     3241 ttttttgata cattcttgga ttctttttgc taatattttg ttgaaaatgt ttgtatcttt
     3301 gttcatgaga gatattggtc tgttgttttc ttttcttgta atgtcatttt ctagttccgg
     3361 tattaaggta atgctggcct agttgaatga tttaggaagt attccctctg cttctgtctt
     3421 ctgaaagaga ttgtagaaag ttgatacaat ttttttttct ttaaatatct tgatagagat
     3481 cttccggact cagatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc
     3541 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     3601 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3661 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3721 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3781 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3841 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3901 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3961 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4021 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4081 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4141 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4201 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4261 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4321 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4381 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4441 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4501 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4561 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4621 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4681 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4741 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4801 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4861 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4921 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     4981 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5041 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5101 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5161 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5221 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5281 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     5341 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5401 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5461 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5521 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5581 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5641 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5701 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     5761 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5821 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5881 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5941 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     6001 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     6061 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6121 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6181 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6241 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6301 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6361 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6421 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6481 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6541 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6601 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6661 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6721 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6781 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6841 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
     6901 tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
