LOCUS       Exported                4736 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 4736)
  AUTHORS   Qinglan Biotech
  TITLE     Synthetic gene from Qinglan Biotech
  JOURNAL   Exported 09-12-2019 from Plasmap http://www.qinglanbiotech.com
REFERENCE   2  (bases 1 to 4736)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 14, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4736
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(37..652)
                     /direction=LEFT
                     /label=pBR322_replication_origin
                     /note="pBR322_replication_origin"
     terminator      660..689
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     primer_bind     724..743
                     /label=Ftw
                     /note="Ftw"
     primer_bind     751..768
                     /label=M13F
                     /note="M13F"
     misc_feature    820..1019
                     /label=14-LTR
     misc_feature    820..1019
                     /label=14 TR
     primer_bind     complement(827..860)
                     /label=Read2-14-L2
     primer_bind     complement(827..860)
                     /label=P7-14-L2
     primer_bind     complement(843..874)
                     /label=14-L1
     misc_feature    851..900
                     /label=2
     misc_feature    851..900
                     /label=1
     misc_feature    953..1002
                     /label=4
     misc_feature    953..1002
                     /label=3
     primer_bind     complement(984..1044)
                     /label=14-NheI-eGFP-IV-R2.6k-IF
     primer_bind     complement(984..1020)
                     /label=14-ASCI-eGFP-IV-F2.6k-IF
     promoter        1035..1534
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             1554..2153
                     /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             2160..2216
                     /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             2217..2936
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2964..3188
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3174..3195
                     /label=ASCI-eGFP-F
     primer_bind     3182..3239
                     /label=14-ASCI-eGFP-IV-F2.6k-IF
     primer_bind     3203..3239
                     /label=14-NheI-eGFP-IV-R2.6k-IF
     misc_feature    3204..3403
                     /label=14-RTR
     misc_feature    complement(3204..3403)
                     /label=14 TR
     misc_feature    complement(3221..3270)
                     /label=4
     misc_feature    complement(3221..3270)
                     /label=3
     misc_feature    complement(3323..3372)
                     /label=2
     misc_feature    complement(3323..3372)
                     /label=1
     primer_bind     3349..3380
                     /label=14-L1
     primer_bind     3363..3396
                     /label=Read2-14-L2
     primer_bind     3363..3396
                     /label=P7-14-L2
     primer_bind     complement(3439..3456)
                     /label=M13R
                     /note="M13R"
     primer_bind     complement(3482..3498)
                     /label=Rtw
                     /note="Rtw"
     terminator      3537..3568
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3569..3671
                     /label=cat promoter
                     /note="cat promoter"
     gene            3672..4532
                     /label=ampicillin_resistance_gene_ORF
                     /note="ampicillin_resistance_gene_ORF"
     CDS             4533..4724
                     /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 tcatatgtac agcggggaaa ataagtattt
      841 gacacatcag catttttatc agtaagggga tttctaagtg ggctactgac acaaaattcc
      901 taccagatgt agccatcaag ccaaatattg aattcataca aagaaatcag aacatttaag
      961 tatacaagtt gagtcataat aaataaagtg aaatgacaca gggaataagt attgaacacg
     1021 ctagcaattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
     1081 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
     1141 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
     1201 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
     1261 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
     1321 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
     1381 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
     1441 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
     1501 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agccaacgcc accatgggga
     1561 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     1621 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     1681 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     1741 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     1801 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     1861 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     1921 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     1981 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     2041 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     2101 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccatcgatg
     2161 ccactaactt ctccctgttg aaacaagcag gggatgtcga agagaatccc gggccaatgg
     2221 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2281 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2341 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2401 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2461 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2521 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2581 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2641 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2701 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2761 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2821 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2881 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaattaa
     2941 ttaagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3001 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3061 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3121 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3181 ctctatggct tctgaggcgc gccgtgttca atacttattc cctgtgtcat ttcactttat
     3241 ttattatgac tcaacttgta tacttaaatg ttctgatttc tttgtatgaa ttcaatattt
     3301 ggcttgatgg ctacatctgg taggaatttt gtgtcagtag cccacttaga aatcccctta
     3361 ctgataaaaa tgctgatgtg tcaaatactt attttccccg ctgtagggcc cagagacgga
     3421 gtcactgcca accgagacgg tcatagctgt ttcctgtgtg ccgcttcctc gctcactgac
     3481 tcgctgcgct cggtcgttac tgaccattta aatcatacct gacctccata gcagaaagtc
     3541 aaaagcctcc gaccggaggc ttttgacttg atcggcacgt aagaggttcc aactttcacc
     3601 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     3661 aggaagctaa aatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3721 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3781 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3841 gtttacgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3901 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3961 agaatgactt ggttgagtac tcaccagtca cagaaaagca tctcacggat ggcatgacag
     4021 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4081 tggcaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4141 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4201 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4261 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggat
     4321 cacttctgcg ctcggccctc ccggctggct ggtttattgc tgataaatct ggagccggtg
     4381 agcgtgggtc ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgcatcg
     4441 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4501 agataggtgc ctcactgatt aagcattggt aagtgaccaa acaggaaaaa accgccctta
     4561 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
     4621 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
     4681 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgatgaggg cccaaa
//
