LOCUS       sequence-58227-e        5672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5672)
  AUTHORS   Ochiai H, Fujita K, Suzuki K, Nishikawa M, Shibata T, Sakamoto N, 
            Yamamoto T
  TITLE     Targeted mutagenesis in the sea urchin embryo using zinc-finger 
            nucleases.
  JOURNAL   Genes Cells. 2010 Aug;15(8):875-85. doi: 
            10.1111/j.1365-2443.2010.01425.x. Epub 2010 Jul 2.
  PUBMED    20604805
REFERENCE   2  (bases 1 to 5672)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1111/j.1365-2443.2010.01425.x"; journalName: "Genes Cells"; 
            date: "2010-08"; volume: "15"; issue: "8"; pages: "875-85"
COMMENT     Sequence Label: pGL4-SSA
FEATURES             Location/Qualifiers
     source          1..5672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      670..679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3226..3347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3237..3256)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3291..3310
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3595..3612)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3766..4354)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3846..3865)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4554..5345,5346..5414))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5519..5567
                     /note="synthetic polyadenylation signal"
     misc_feature    5581..5672
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5621..5640
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct 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 cagagctggt ttagtgaacc
      601 gtcagatccg ctagcgctac cggactcaga tctcgagctc aagcttggca atccggtact
      661 gttggtaaag ccaccatgga agatgccaaa aacattaaga agggcccagc gccattctac
      721 ccactcgaag acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg ctacgccctg
      781 gtgcccggca ccatcgcctt taccgacgca catatcgagg tggacattac ctacgccgag
      841 tacttcgaga tgagcgttcg gctggcagaa gctatgaagc gctatgggct gaatacaaac
      901 catcggatcg tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt gttgggtgcc
      961 ctgttcatcg gtgtggctgt ggccccagct aacgacatct acaacgagcg cgagctgctg
     1021 aacagcatgg gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg gctgcaaaag
     1081 atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat ggatagcaag
     1141 accgactacc agggcttcca aagcatgtac accttcgtga cttcccattt gccacccggc
     1201 ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg acaaaaccat cgccctgatc
     1261 atgaacagta gtggcagtac cggattgccc aagggcgtag ccctaccgca ccgcaccgct
     1321 tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca accagatcat ccccgacacc
     1381 gctatcctca gcgtggtgcc atttcaccac ggcttcggca tgttcaccac gctgggctac
     1441 ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg aggaggagct attcttgcgc
     1501 agcttgcaag actataagat tcaatctgcc ctgctggtgc ccacactatt tagcttcttc
     1561 gctaagagca ctctcatcga caagtacgac ctaagcaact tgcacgagat cgccagcggc
     1621 ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca aacgcttcca cctaccaggc
     1681 atccgccagg gctacggcct gacagaaaca accagcgcca ttctgatcac ccccgaaggg
     1741 gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa ggtggtggac
     1801 ttggactagg gtctctgtcg tgcccgggta ctgatgtacc gtgagaccta ggagcgcgag
     1861 ctgctgaaca gcatgggcat cagccagccc accgtcgtat tcgtgagcaa gaaagggctg
     1921 caaaagatcc tcaacgtgca aaagaagcta ccgatcatac aaaagatcat catcatggat
     1981 agcaagaccg actaccaggg cttccaaagc atgtacacct tcgtgacttc ccatttgcca
     2041 cccggcttca acgagtacga cttcgtgccc gagagcttcg accgggacaa aaccatcgcc
     2101 ctgatcatga acagtagtgg cagtaccgga ttgcccaagg gcgtagccct accgcaccgc
     2161 accgcttgtg tccgattcag tcatgcccgc gaccccatct tcggcaacca gatcatcccc
     2221 gacaccgcta tcctcagcgt ggtgccattt caccacggct tcggcatgtt caccacgctg
     2281 ggctacttga tctgcggctt tcgggtcgtg ctcatgtacc gcttcgagga ggagctattc
     2341 ttgcgcagct tgcaagacta taagattcaa tctgccctgc tggtgcccac actatttagc
     2401 ttcttcgcta agagcactct catcgacaag tacgacctaa gcaacttgca cgagatcgcc
     2461 agcggcgggg cgccgctcag caaggaggta ggtgaggccg tggccaaacg cttccaccta
     2521 ccaggcatcc gccagggcta cggcctgaca gaaacaacca gcgccattct gatcaccccc
     2581 gaaggggacg acaagcctgg cgcagtaggc aaggtggtgc ccttcttcga ggctaaggtg
     2641 gtggacttgg acaccggtaa gacactgggt gtgaaccagc gcggcgagct gtgcgtccgt
     2701 ggccccatga tcatgagcgg ctacgttaac aaccccgagg ctacaaacgc tctcatcgac
     2761 aaggacggct ggctgcacag cggcgacatc gcctactggg acgaggacga gcacttcttc
     2821 atcgtggacc ggctgaagag cctgatcaaa tacaagggct accaggtagc cccagccgaa
     2881 ctggagagca tcctgctgca acaccccaac atcttcgacg ccggggtcgc cggcctgccc
     2941 gacgacgatg ccggcgagct gcccgccgca gtcgtcgtgc tggaacacgg taaaaccatg
     3001 accgagaagg agatcgtgga ctatgtggcc agccaggtta caaccgccaa gaagctgcgc
     3061 ggtggtgttg tgttcgtgga cgaggtgcct aaaggactga ccggcaagtt ggacgcccgc
     3121 aagatccgcg agattctcat taaggccaag aagggcggca agatcgccgt gaattcttaa
     3181 ctgcagtcta gagtcggggc ggccggccgc ttcgagcaga catgataaga tacattgatg
     3241 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3301 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3361 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     3421 acctctacaa atgtggtaaa atcgataagg atccgtcgac cgatgccctt gagagccttc
     3481 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     3541 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct
     3601 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3661 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3721 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3781 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3841 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3901 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3961 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4021 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4081 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4141 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4201 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4261 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4321 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4381 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4441 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4501 atatgagtaa acttggtctg acagcggccg caaatgctaa accactgcag tggttaccag
     4561 tgcttgatca gtgaggcacc gatctcagcg atctgcctat ttcgttcgtc catagtggcc
     4621 tgactccccg tcgtgtagat cactacgatt cgtgagggct taccatcagg ccccagcgca
     4681 gcaatgatgc cgcgagagcc gcgttcaccg gcccccgatt tgtcagcaat gaaccagcca
     4741 gcagggaggg ccgagcgaag aagtggtcct gctactttgt ccgcctccat ccagtctatg
     4801 agctgctgtc gtgatgctag agtaagaagt tcgccagtga gtagtttccg aagagttgtg
     4861 gccattgcta ctggcatcgt ggtatcacgc tcgtcgttcg gtatggcttc gttcaactct
     4921 ggttcccagc ggtcaagccg ggtcacatga tcacccatat tatgaagaaa tgcagtcagc
     4981 tccttagggc ctccgatcgt tgtcagaagt aagttggccg cggtgttgtc gctcatggta
     5041 atggcagcac tacacaattc tcttaccgtc atgccatccg taagatgctt ttccgtgacc
     5101 ggcgagtact caaccaagtc gttttgtgag tagtgtatac ggcgaccaag ctgctcttgc
     5161 ccggcgtcta tacgggacaa caccgcgcca catagcagta ctttgaaagt gctcatcatc
     5221 gggaatcgtt cttcggggcg gaaagactca aggatcttgc cgctattgag atccagttcg
     5281 atatagccca ctcttgcacc cagttgatct tcagcatctt ttactttcac cagcgtttcg
     5341 gggtgtgcaa aaacaggcaa gcaaaatgcc gcaaagaagg gaatgagtgc gacacgaaaa
     5401 tgttggatgc tcatactcgt cctttttcaa tattattgaa gcatttatca gggttactag
     5461 tacgtctctc aaggataagt aagtaatatt aaggtacggg aggtattgga caggccgcaa
     5521 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact
     5581 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
     5641 cagtgcaagt gcaggtgcca gaacatttct ct
//
