LOCUS       40924_10276        5759 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5759)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5759)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        632..650
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    680..704
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             713..1429
                     /codon_start=1
                     /label=EmGFP
                     /note="Emerald GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     ncRNA           1492..1518
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
     ncRNA           1582..1623
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse
                     miR-155 microRNA (Uva et al., 2013)"
     protein_bind    complement(1652..1676)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1762..1810
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(1981..1999)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2004..2020)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2088..2516
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2530..2859
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        2907..2954
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2973..3368
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3529..3662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3856..4444)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4543..5331)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      601 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagtcccaag
      661 ctggctagtt aagctatcaa caagtttgta caaaaaagca ggctttaaaa ccatggtgag
      721 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      781 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      841 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      901 caccttcacc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
      961 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1021 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1081 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1141 gtacaactac aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa
     1201 ggtgaacttc aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1261 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1321 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1381 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagctaagca
     1441 cttcgtggcc gtcgatcgtt taaagggagg tagtgagtcg accagtggat cctggaggct
     1501 tgctgaaggc tgtatgctga aatgtactgc gcgtggagac gttttggcca ctgactgacg
     1561 tctccacgca gtacatttca ggacacaagg cctgttacta gcactcacat ggaacaaatg
     1621 gcccagatct ggccgcactc gagatatcta gacccagctt tcttgtacaa agtggttgat
     1681 ctagagggcc cgcggttcgc tgatggggga ggctaactga aacacggaag gagacaatac
     1741 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     1801 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     1861 tgaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     1921 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcatcc
     1981 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
     2041 aaaatctctg atggatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     2101 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2161 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2221 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2281 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2341 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2401 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2461 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2521 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2581 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2641 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2701 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2761 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2821 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2881 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     2941 tagtataata cgacaaggtg aggaactaaa ccatggccaa gcctttgtct caagaagaat
     3001 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     3061 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     3121 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     3181 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     3241 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     3301 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     3361 gggagggcta agcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     3421 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3481 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     3541 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3601 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     3661 ataccgtcgc tcttccgctg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3721 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3781 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3841 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3901 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3961 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4021 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4081 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4141 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4201 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4261 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4321 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4381 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4441 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgtaact
     4501 cacgttaagg gattttggtc atgggtggct cgacgagggt tatttgccga ctaccttggt
     4561 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
     4621 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
     4681 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     4741 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
     4801 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     4861 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     4921 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     4981 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     5041 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     5101 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     5161 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     5221 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     5281 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca taatgtttaa
     5341 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata acatcaaaca
     5401 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtaccccaaa
     5461 aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt
     5521 caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagct tacgaaccga
     5581 acaggcttat gtccactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc
     5641 aaccttgggt agcagcgaag tcgaggcatt tctgtcctgg ctggtctaga attgcatgaa
     5701 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgc
//
