LOCUS       Exported                4768 bp ds-DNA     circular SYN 13-SEP-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4768)
  AUTHORS   Qinglan Biotech
  TITLE     Synthetic gene from Qinglan Biotech
  JOURNAL   Exported 19-11-2019 from Plasmap http://www.qinglanbiotech.com
REFERENCE   2  (bases 1 to 4768)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 14, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4768
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(37..652)
                     /direction=LEFT
                     /label=pBR322_replication_origin
                     /note="pBR322_replication_origin"
     primer_bind     724..743
                     /label=Ftw
                     /note="Ftw"
     primer_bind     751..768
                     /label=M13F
                     /note="M13F"
     misc_feature    820..1036
                     /label=36LTR
     misc_feature    820..1034
                     /label=36 TR(mismatch)
     misc_feature    820..882
                     /label=36 TR
     misc_feature    820..848
                     /label=3
     misc_feature    820..848
                     /label=3
     misc_feature    820..848
                     /label=2
     misc_feature    820..848
                     /label=2
     primer_bind     complement(827..852)
                     /label=Read2-36-L2
     primer_bind     complement(827..852)
                     /label=P7-36-L2
     primer_bind     complement(841..868)
                     /label=36-L1
     misc_feature    1008..1036
                     /label=4
     primer_bind     complement(1012..1034)
                     /label=36-ASCI-eGFP-IV-F2.6k-IF
     primer_bind     complement(1015..1061)
                     /label=36-NheI-eGFP-IV-R2.6k-IF
     promoter        1052..1551
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1322..1333)
                     /label=T7ter
                     /note="T7ter"
     CDS             1571..2170
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             2177..2233
                     /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"
     CDS             2234..2953
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2981..3205
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3199..3245
                     /label=36-ASCI-eGFP-IV-F2.6k-IF
     misc_feature    3213..3220
                     /locus_tag="MCS(5)"
                     /label=MCS(5)
     misc_feature    3221..3435
                     /label=36RTR
     misc_feature    complement(3221..3249)
                     /label=1
     misc_feature    complement(3223..3435)
                     /label=36 TR
     primer_bind     3223..3242
                     /label=36-NheI-eGFP-IV-R2.6k-IF
     misc_feature    complement(3373..3435)
                     /label=36 TR
     primer_bind     3387..3414
                     /label=36-L1
     primer_bind     3403..3428
                     /label=Read2-36-L2
     primer_bind     3403..3428
                     /label=P7-36-L2
     misc_feature    complement(3407..3435)
                     /label=3
     misc_feature    complement(3407..3435)
                     /label=2
     primer_bind     complement(3471..3488)
                     /label=M13R
                     /note="M13R"
     primer_bind     complement(3514..3530)
                     /label=Rtw
                     /note="Rtw"
     terminator      3569..3600
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3601..3703
                     /label=cat promoter
                     /note="cat promoter"
     gene            3704..4564
                     /label=ampicillin_resistance_gene_ORF
                     /note="ampicillin_resistance_gene_ORF"
     CDS             4565..4756
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg
       61 ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg
      121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      481 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac ctcggaaaaa
      541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg attttctacc
      661 gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgcagtccag ttacgctgga
      721 gtcaataggc gtatcacgag gccctttcgt tgtaaaacga cggccagtcg aaccacgcaa
      781 tgcgtctcga tccgcagtgt cttgcgtctc taagctttac agtggtgtga aaaactattt
      841 gcccccttcc tgatttctta ttcttttgca tgtttgtcac acttaaatgt ttctgctcat
      901 caaaaaccgt taactattag tcaaagataa cataattgaa cacaaaatgc agtttttaaa
      961 tgaaggttta cgttattaag ggagaaaaaa aactccaaat ctacatggcc ctgtgtgaaa
     1021 aagtgattgc ccccctgcta gcaattctac cgggtagggg aggcgctttt cccaaggcag
     1081 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
     1141 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
     1201 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
     1261 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
     1321 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
     1381 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
     1441 ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa
     1501 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc
     1561 caacgccacc atggggaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     1621 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     1681 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     1741 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     1801 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     1861 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     1921 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     1981 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     2041 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     2101 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     2161 gcccggtgcc atcgatgcca ctaacttctc cctgttgaaa caagcagggg atgtcgaaga
     2221 gaatcccggg ccaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2281 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2341 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2401 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2461 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2521 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2581 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2641 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2701 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2761 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2821 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2881 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2941 gctgtacaag taattaatta agctcgctga tcagcctcga ctgtgccttc tagttgccag
     3001 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3061 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3121 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3181 gctggggatg cggtgggctc tatggcttct gaggcgcgcc cagggggcaa ttactttttc
     3241 acacagggcc atgtaggttt ggattttttt tctccctaaa taataaaaac cctcatttaa
     3301 aaactgcatt ttgtgtttac ttgtgttatc tttgactaat agttaaatgt gtttgatgat
     3361 cagaaacatt ttgtgtgaca aacatgcaaa agaataagaa atcaggaagg gggcaaatag
     3421 tttttcacac cactgtaggg cccagagacg gagtcactgc caaccgagac ggtcatagct
     3481 gtttcctgtg tgccgcttcc tcgctcactg actcgctgcg ctcggtcgtt actgaccatt
     3541 taaatcatac ctgacctcca tagcagaaag tcaaaagcct ccgaccggag gcttttgact
     3601 tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
     3661 attttttgag ttatcgagat tttcaggagc taaggaagct aaaatgagta ttcaacattt
     3721 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3781 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3841 actggatctc aacagcggta agatccttga gagtttacgc cccgaagaac gttttccaat
     3901 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3961 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4021 cacagaaaag catctcacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4081 catgagtgat aacactgcgg ccaacttact tctggcaacg atcggaggac cgaaggagct
     4141 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4201 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4261 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4321 agactggatg gaggcggata aagttgcagg atcacttctg cgctcggccc tcccggctgg
     4381 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcccgcggta tcattgcagc
     4441 actggggcca gatggtaagc cctcccgcat cgtagttatc tacacgacgg ggagtcaggc
     4501 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4561 gtaagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
     4621 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
     4681 atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga
     4741 cggtgaaaac ctctgatgag ggcccaaa
//
