LOCUS       sequence-141619-        4256 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 4256)
  AUTHORS   Kim JH, Lee SR, Li LH, Park HJ, Park JH, Lee KY, Kim MK, Shin BA, 
            Choi SY
  TITLE     High Cleavage Efficiency of a 2A Peptide Derived from Porcine 
            Teschovirus-1 in Human Cell Lines, Zebrafish and Mice.
  JOURNAL   PLoS One. 2011  . 6(4):e18556.
  PUBMED    21602908
REFERENCE   2  (bases 1 to 4256)
  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"; journalName: "PLoS One. 
            2011  . 6(4):e18556."
COMMENT     Sequence Label: pSYC-181
FEATURES             Location/Qualifiers
     source          1..4256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             180..236
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(281..300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(282..300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    305..439
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(329..348)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     383..402
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(552..572)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(552..570)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(591..607)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(591..607)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(604..626)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    615..631
                     /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(join(639..645,646..663,664..669))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    684..705
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(822..839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(993..1581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1073..1092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1752..2543,2544..2612))
                     /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"
     primer_bind     2375..2394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2613..2717)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2743..3198)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2880..2901)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3092..3111
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3272..4256
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttgtacagat atctggcgcg cctaccatgg
      121 ggcccggccg gcctaagatc accggtcccg ggcttaagga gctcgcatgc ggaagcggag
      181 ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctagat
      241 ctggaattca aggcctctcg agcctctaga tagctagcct ccctatagtg agtcgtatta
      301 cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
      361 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
      421 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
      481 ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc
      541 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
      601 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata
      661 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      721 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      781 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      841 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      901 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      961 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1021 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1081 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1141 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1201 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1261 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1321 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1381 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     1441 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1501 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1561 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1621 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1681 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1741 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1801 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1861 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1921 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1981 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2041 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2101 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2161 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2221 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2281 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2341 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2401 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2461 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2521 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2581 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2641 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2701 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     2761 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2821 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2881 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2941 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3001 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3061 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3121 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3181 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     3241 cggtgcgggc ctcttcgcta ttacgccagt cgaccatagc caattcaata tggcgtatat
     3301 ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat
     3361 ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata tggcggactt
     3421 ggcaccatgc caattcaata tggcggactt ggcactgtgc caactgggga ggggtctact
     3481 tggcacggtg ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat
     3541 tggcatggtg ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa
     3601 tatccaatat ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc
     3661 ctatgccata tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat
     3721 gggcggtccc atataccata tatggggctt cctaataccg cccatagcca ctcccccatt
     3781 gacgtcaatg gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga
     3841 cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc
     3901 ccattgacgt caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc
     3961 atatccgttc tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac
     4021 atcaatatct attaatagta acttggcaag tacattacta ttggaagtac gccagggtac
     4081 attggcagta ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac
     4141 atccccattg acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa
     4201 tgggcggtag gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaac
//
