LOCUS       Exported                8656 bp DNA     circular SYN 19-NOV-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8656)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8656)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8656)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     CDS             1764..1784
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1806..1826
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1839..2027
                     /codon_start=1
                     /label=zinc finger domain
                     /translation="GNTSGVLSTPKAKRAKHPPGTEKPRSRSQSEQPATCPICYAVIRQ
                     SRNLRRHLELRHFAKPGV"
     CDS             2028..2765
                     /codon_start=1
                     /label=SceGFP
                     /translation="DPPVATMVSKGEELFTGVVPILVELDGDVNGHKFSVSG*G*QGNT
                     YGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERT
                     IFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADK
                     QKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKR
                     DHMVLLEFVTAAGITLGMDELYK"
     polyA_signal    2888..3009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3670..4269)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(4288..4785)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     polyA_signal    4938..4993
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     CDS             5357..5377
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5399..5419
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5432..5620
                     /codon_start=1
                     /label=zinc finger domain
                     /translation="GNTSGVLSTPKAKRAKHPPGTEKPRSRSQSEQPATCPICYAVIRQ
                     SRNLRRHLELRHFAKPGV"
     CDS             5621..6153
                     /codon_start=1
                     /label=iGFP
                     /translation="DPPVATMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDAT
                     YGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERT
                     IFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADK
                     QKNGIR*TSIKAWR"
     regulatory      5633..5642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6159..6175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6183..6199
                     /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(6207..6237)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6252..6273
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6332..6527
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6378..6513
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6533..6667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6906..7494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7665..8525)
                     /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(8526..8630)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcagatccg
     1741 ccgccactat gggatcaaga tcgccaaaaa agaagagaaa ggtgccgaag aagcatgcag
     1801 caccaccaaa aaaaaaacga aaagtagaag acccacgagg caacaccagc ggcgtgctga
     1861 gcacccccaa ggccaagagg gccaagcacc cccccggcac cgagaagccc aggagcagga
     1921 gccagagcga gcagcccgcc acctgcccca tctgctacgc cgtgatcagg cagagcagga
     1981 acctgaggag gcacctggag ctgaggcact tcgccaagcc cggcgtggat ccaccggtcg
     2041 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2101 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg ctagggataa cagggtaata
     2161 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2221 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2281 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2341 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2401 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2461 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     2521 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     2581 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2641 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     2701 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2761 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     2821 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2881 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2941 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3001 tgtatcttaa ggcgtaaatt gtaagcgtta atctgaagct tctgatggaa ttagaacttg
     3061 gcaaaacaat actgagaatg aagtgtatgt ggaacagagg ctgctgatct cgttcttcag
     3121 gctatgaaac tgacacattt ggaaaccaca gtacttagaa ccacaaagtg ggaatcaaga
     3181 gaaaaacaat gatcccacga gagatctata gatctataga tcatgagtgg gaggaatgag
     3241 ctggccctta atttggtttt gcttgtttaa attatgatat ccaactatga aacattatca
     3301 taaagcaata gtaaagagcc ttcagtaaag agcaggcatt tatctaatcc caccccaccc
     3361 ccacccccgt agctccaatc cttccattca aaatgtaggt actctgttct cacccttctt
     3421 aacaaagtat gacaggaaaa acttccattt tagtggacat ctttattgtt taatagatca
     3481 tcaatttcga tccgctcctg ggcaccgaac tgcgccgcgt gttcagcagg gtcggcgtgt
     3541 tcggtgtgtc ccccgcggtg ggcctcgggg gcgggtgcgg ggtcggcggg gccgccccgg
     3601 gtggcttcgg tcggagccat ggggtcgtgc gctcctttcg gtcgggcgct gcgggtcgtg
     3661 gggcgggcgt caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac
     3721 ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc
     3781 ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc cggggagcac
     3841 gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg tggccaggaa
     3901 ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt gctgcgcggc
     3961 cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc
     4021 ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg cgacccacac
     4081 cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc
     4141 gatgtggcgg tccgggtcga cggtgtggcg cgtggcgggg tagtcggcga acgcggcggc
     4201 gagggtgcgt acggcccggg ggacgtcgtc gcgggtggcg aggcgcaccg tgggcttgta
     4261 ctcggtcatg gtggcggctg gatcggtcga aaggcccgga gatgaggaag aggagaacag
     4321 cgcggcagac gtgcgctttt gaagcgtgca gaatgccggg cctccggagg accttcgggc
     4381 gcccgccccg cccctgagcc cgcccctgag cccgcccccg gacccacccc ttcccagcct
     4441 ctgagcccag aaagcgaagg agcaaagctg ctattggccg ctgccccaaa ggcctacccg
     4501 cttccattgc tcagcggtgc tgtccatctg cacgagacta gtgagacgtg ctacttccat
     4561 ttgtcacgtc ctgcacgacg cgagctgcgg ggcggggggg aacttcctga ctaggggagg
     4621 agtagaaggt ggcgcgaagg ggccaccaaa gaacggagcc ggttggcgcc taccggtgga
     4681 tgtggaatgt gtgaggccag aggccacttg tgtagcgcca agtgcccagc ggggctgcta
     4741 aagcgcatgc tccagactgc cttgggaaaa gcgcctcccc tacccggtag aattcactcc
     4801 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     4861 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     4921 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     4981 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     5041 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     5101 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     5161 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     5221 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     5281 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
     5341 ttagggatca agatcgccaa aaaagaagag aaaggtgccg aagaagcatg cagcaccacc
     5401 aaaaaaaaaa cgaaaagtag aagacccacg aggcaacacc agcggcgtgc tgagcacccc
     5461 caaggccaag agggccaagc acccccccgg caccgagaag cccaggagca ggagccagag
     5521 cgagcagccc gccacctgcc ccatctgcta cgccgtgatc aggcagagca ggaacctgag
     5581 gaggcacctg gagctgaggc acttcgccaa gcccggcgtg gatccaccgg tcgccaccat
     5641 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     5701 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     5761 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     5821 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     5881 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     5941 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     6001 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     6061 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     6121 catcaggtga acttcaataa aagcttggcg taatcatggt catagctgtt tcctgtgtga
     6181 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6241 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6301 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc
     6361 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6421 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     6481 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt
     6541 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6601 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6661 ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6721 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6781 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6841 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6901 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6961 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7021 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7081 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7141 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7201 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7261 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7321 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7381 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7441 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7501 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7561 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7621 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7681 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7741 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7801 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7861 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7921 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7981 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     8041 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     8101 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     8161 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     8221 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     8281 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8341 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8401 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     8461 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8521 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8581 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8641 cgaaaagtgc cacctg
//
