LOCUS       Exported                8543 bp DNA     circular SYN 14-NOV-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    Mega PiggyBac Transposase
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8543)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8543)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8543)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8543)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          3411..3422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..1993
                     /label=chicken 5'-HS4 beta-globin insulator (HS4)
                     /note="Insulators are DNA sequence elements that prevent 
                     inappropriate interactions between adjacent chromatin 
                     domains. One type of insulator establishes domains that 
                     separate enhancers and promoters to block their 
                     interaction, whereas a second type creates a barrier 
                     against the spread of heterochromatin."
     primer_bind     2449..2465
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2492..3197
                     /label=Polr2A promoter
                     /note="RNA polymerase II large subunit promoter"
     intron          3454..3550
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             3626..5407
                     /codon_start=1
                     /product="Mega-PiggyBac"
                     /label=Mega-PiggyBac
                     /note="Mega-PiggyBac"
                     /translation="MGSSLDDEHILSALLQSDDELVGEDSDSEVSDHVSEDDVQSDTEE
                     AFIDEVHEVQPTSSGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKPT
                     RRSRVSALNIVRSQRGPTRMCRNIYDPLLCFKLFFTDEIISEIVKWTNAEISLKRRESM
                     TSATFRDTNEDEIRAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIR
                     CLRMDDKSIRPTLRESDVFTPVRKIWDIFINQCIQNYTPGAHLTIDEQLLGFRGRCPFR
                     VYIPNKPSKYGIKIVMICDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVRGSC
                     RNITCDNWFTSIPLAKNLLQEPYKLTLVGTVRSNKREIPEELKNSRSRPVGTSMFCFDG
                     PLTLVSYKPKPAKMVYLLSSCDEDAIINETTGKPEMIMYYNQTKGGVDTLDQMCSLMSC
                     SRKTNRWPMALFYGMINIACINSYIIYCHNVSSKGEKVQSRKKFMKNLYKGLTSSFMRK
                     RLEAPTLKRYLRDNISNILPKEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKASASCKK
                     CKKVICREHNIDMCQSCF"
     polyA_signal    complement(5436..5570)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(5687..5705)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5726..5742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5750..5766)
                     /label=lac operator
                     /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(5774..5804)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5819..5840)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6128..6716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6890..7747)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7748..7852)
                     /label=AmpR promoter
     rep_origin      7879..8334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8475..8491
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8501..8519
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcggccgcga gctcacgggg acagcccccc cccaaagccc ccagggatgt aattacgtcc
       61 ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc
      121 ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga
      181 tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg
      241 gggaaaaagc tttaggctga aagagagatt tagaatgaca gaatcataga acggcctggg
      301 ttgcaaagga gcacagtgct catccagatc caaccccctg ctatgtgcag ggtcatcaac
      361 cagcagccca ggctgcccag agccacatcc agcctggcct tgaatgcctg cagggatggg
      421 gcatccacag cctccttggg caacctgttc agtgcgtcac caccctctgg gggaaaaact
      481 gcctcctcat atccaaccca aacctcccct gtctcagtgt aaagccattc ccccttgtcc
      541 tatcaagggg gagtttgctg tgacattgtt ggtctggggt gacacatgtt tgccaattca
      601 gtgcatcacg gagaggcaga tcttggggat aaggaagtgc aggacagcat ggacgtggga
      661 catgcaggtg ttgagggctc tgggacactc tccaagtcac agcgttcaga acagccttaa
      721 ggataagaag ataggataga aggacaaaga gcaagttaaa acccagcatg gagaggagca
      781 caaaaaggcc acagacactg ctggtccctg tgtctgagcc tgcatgtttg atggtgtctg
      841 gatgcaagca gaaggggtgg aagagcttgc ctggagagat acagctgggt cagtaggact
      901 gggacaggca gctggagaat tgccatgtag atgttcatac aatcgtcaaa tcatgaaggc
      961 tggaaaagcc ctccaagatc cccaagacca accccaaccc acccaccgtg cccactggcc
     1021 atgtccctca gtgccacatc cccacagttc ttcatcacct ccagggacgg tgaccccccc
     1081 acctccgtgg gcagctgtgc cactgcagca ccgctctttg gagaaggtaa atcttgctaa
     1141 atccagcccg accctcccct ggcacaacgt aaggccatta tctctcatcc aactccagga
     1201 cggagtcagt gagaatattg cctccctggc gagctcacgg ggacagcccc cccccaaagc
     1261 ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc gcccggggct
     1321 ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg
     1381 gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc
     1441 tgcagacacc tggggggata cggggaaaaa gctttaggct gaaagagaga tttagaatga
     1501 cagaatcata gaacggcctg ggttgcaaag gagcacagtg ctcatccaga tccaaccccc
     1561 tgctatgtgc agggtcatca accagcagcc caggctgccc agagccacat ccagcctggc
     1621 cttgaatgcc tgcagggatg gggcatccac agcctccttg ggcaacctgt tcagtgcgtc
     1681 accaccctct gggggaaaaa ctgcctcctc atatccaacc caaacctccc ctgtctcagt
     1741 gtaaagccat tcccccttgt cctatcaagg gggagtttgc tgtgacattg ttggtctggg
     1801 gtgacacatg tttgccaatt cagtgcatca cggagaggca gatcttgggg ataaggaagt
     1861 gcaggacagc atggacgtgg gacatgcagg tgttgagggc tctgggacac tctccaagtc
     1921 acagcgttca gaacagcctt aaggataaga agataggata gaaggacaaa gagcaagtta
     1981 aaacccagca tggagaggag cacaaaaagg ccacagacac tgctggtccc tgtgtctgag
     2041 cctgcatgtt tgatggtgtc tggatgcaag cagaaggggt ggaagagctt gcctggagag
     2101 atacagctgg gtcagtagga ctgggacagg cagctggaga attgccatgt agatgttcat
     2161 acaatcgtca aatcatgaag gctggaaaag ccctccaaga tccccaagac caaccccaac
     2221 ccacccaccg tgcccactgg ccatgtccct cagtgccaca tccccacagt tcttcatcac
     2281 ctccagggac ggtgaccccc ccacctccgt gggcagctgt gccactgcag caccgctctt
     2341 tggagaaggt aaatcttgct aaatccagcc cgaccctccc ctggcacaac gtaaggccat
     2401 tatctctcat ccaactccag gacggagtca gtgagaatat tcggccgctc tagaactagt
     2461 ggatcccccg ggctgcagga attcgatggc cgcttgggcg ggatatctag cgccaaggtt
     2521 ccgtaaagct ctctattaga ggactggcgg tgccccatca ccctgggagg cctctccaca
     2581 aatatcggct acttccaatt gattggacgc gccatcttgt ctgcttatgc atattcagag
     2641 gatcctgaat attcatgagt gagggccgtg cggcccctcc ctccaaccct cccccggaac
     2701 aagctccgga gaacccgaca ggccccgcct tctttaccga tgcgtagaac aaaccatttt
     2761 ccgggttggg gggggaaatt aatgagagac tttagctgaa aatgagcatg gacgccaaag
     2821 ctgagtaaag atggcttaac tttatcctcc attctgtaat ccgtcagctt gagtgtacgg
     2881 gaagtcggcg aggggggcgg caggctcaga aacattctcc tcctcctgtc gcgtcagaaa
     2941 gaacacccaa ccagggagcc ggagccctgg cgtcaacatc tctggcgcgc gcgctccatg
     3001 taggccggtg cgggcggccc cgtagcgcaa gggagggcgg gaaaggaagg ggcgggacac
     3061 aagggcgaat ctataaaggg cgtcattcag ccagttctct cctcagaagc gccgagagcg
     3121 cgaccgggac ggttggagaa gaaggtggct cccggaaggg ggagagacaa actgccgtaa
     3181 cctctgccgt tcaggatccc ggccatcaag cttatcgatg ctgccgcagc aaaagcagga
     3241 gcagatgccg ccgtcgcagg cgaagatgtc gcagacggag gaggcgatgc tgccggcgga
     3301 ggaggcgaag taagtagagg gctgggctgg gctgtggggg gtgtggggtg cgggactggg
     3361 cagtctggga gtccctctca ccacttttct tacctttcta ggatgctgcc gtcgacgccg
     3421 cttcgaggaa ctgaaaaacc agaaagttaa ctggtaagtt tagtcttttt gtcttttatt
     3481 tcaggtcccg gatccggtgg tggtgcaaat caaagaactg ctcctcagtg gatgttgcct
     3541 ttacttctag gcctgtacgg aagtgttact tctgctctaa aagctgcgga attgtaccca
     3601 attcgttaag gccaaattgg ccaccatggg gagcagtctc gacgatgaac atatactttc
     3661 agccctcctc cagagtgacg acgaactcgt aggcgaagac agtgatagtg aagtctccga
     3721 tcacgtctct gaagatgacg tgcagtcaga cacagaagaa gccttcatcg acgaagtgca
     3781 tgaggtccag cccaccagct cagggtccga gattctggac gagcaaaacg tgatcgagca
     3841 gcctggatca tctctggcaa gtaatcgaat cctcaccctg ccacagagaa cgatccgggg
     3901 aaagaacaaa cattgctgga gtacatctaa gccaactaga aggagccgcg tgagcgctct
     3961 taacatcgtg aggtcacaga ggggcccgac gcgaatgtgc agaaacatct atgatccctt
     4021 gctgtgtttc aagctgtttt tcacagatga gatcataagc gaaatcgtca aatggacaaa
     4081 tgcggaaatt tctctgaaaa ggcgagagag tatgaccagc gcaacatttc gcgatacaaa
     4141 cgaagatgaa atccgggcat tctttggaat tcttgtcatg acagctgttc ggaaggataa
     4201 ccatatgtcc actgacgacc tgttcgaccg gtctctcagc atggtgtatg tgagcgtgat
     4261 gagcagggac agattcgact tcctcattag atgtctccga atggacgaca aatccatccg
     4321 gcccactctc agagaaagcg acgtatttac ccctgtcaga aaaatctggg atatctttat
     4381 aaaccaatgt atccagaatt acacaccagg tgctcatctt acaatcgacg agcaactcct
     4441 gggatttagg gggcgctgtc catttcgagt gtacattccg aataagcctt caaaatacgg
     4501 gatcaagatc gtcatgatct gtgatagtgg gactaaatac atgatcaacg ggatgcccta
     4561 cctgggtaga gggacgcaga ccaatggggt cccactcgga gagtattacg tgaaggagct
     4621 gagcaagccg gtgcggggat cctgtagaaa cattacatgt gataactggt ttacctctat
     4681 cccgctcgcg aagaaccttc tgcaagaacc ttacaagctc acattggttg gcaccgtgcg
     4741 gtccaacaaa cgcgagatcc ccgaggagtt gaagaactca cgctccaggc ccgtaggcac
     4801 cagtatgttc tgcttcgacg ggccacttac tctcgtgagt tataagccta agcccgccaa
     4861 gatggtctac ttgctttcaa gctgcgacga ggacgccatt attaacgaga ccaccggcaa
     4921 gcctgagatg ataatgtatt acaaccagac taagggaggc gtggacacac tggaccagat
     4981 gtgctccctt atgagttgtt caagaaagac caaccggtgg ccaatggcac tcttctacgg
     5041 aatgattaac atcgcctgca ttaacagtta catcatctac tgtcataacg tctcatcaaa
     5101 gggagaaaaa gtacagagca ggaaaaaatt catgaaaaac ctgtacaaag ggctgacatc
     5161 atctttcatg cggaaaaggc tggaggcgcc aacattgaag cggtatctga gagacaacat
     5221 ctccaacatc cttccaaaag aggttcctgg cactagtgat gattcaacag aagagcccgt
     5281 gatgaagaaa cggacatact gcacttattg ccccagcaag atcagaagga aggcgtccgc
     5341 atcctgcaaa aaatgtaaga aagtgatctg cagggagcat aacattgaca tgtgtcagtc
     5401 ttgtttttga ggccgtaacg gccgccagaa ttggggatcc agacatgata agatacattg
     5461 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     5521 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     5581 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt tcggatcctc
     5641 tagagtcgac ctgcaggcat gcaagctcgg tacccagctt ttgttccctt tagtgagggt
     5701 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5761 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5821 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5881 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5941 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6001 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6061 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6121 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6181 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6241 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6301 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6361 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6421 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6481 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6541 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6601 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6661 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6721 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6781 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6841 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6901 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6961 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7021 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7081 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7141 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7201 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7261 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7321 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7381 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7441 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7501 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7561 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7621 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7681 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7741 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7801 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7861 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     7921 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     7981 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     8041 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     8101 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     8161 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     8221 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     8281 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc
     8341 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     8401 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     8461 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     8521 cgaattggag ctccaccgcg gtg
//
