LOCUS       Exported                4861 bp ds-DNA     circular SYN 09-MAR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4861)
  AUTHORS   Qinglan Biotech
  TITLE     Synthetic gene from Qinglan Biotech
  JOURNAL   Exported 22-11-2019 from Plasmap http://www.qinglanbiotech.com
REFERENCE   2  (bases 1 to 4861)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 14, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4861
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      12..39
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     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"
     primer_bind     752..768
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    820..1081
                     /label=35 TR(mismatch)
     misc_feature    820..1081
                     /label=35C LTR
     misc_feature    820..837
                     /label=35 TR
     misc_feature    821..862
                     /label=3
     primer_bind     complement(828..856)
                     /label=Read2-35-L2
     primer_bind     complement(828..856)
                     /label=P7-35-L2
     primer_bind     complement(837..879)
                     /label=35-L1
     misc_feature    1016..1057
                     /label=2
     primer_bind     complement(1037..1106)
                     /label=35-NheI-eGFP-IV-R2.6k-IF
     promoter        1097..1596
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1367..1378)
                     /label=T7ter
                     /note="T7ter"
     CDS             1616..2215
                     /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             2222..2278
                     /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             2279..2998
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3026..3250
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3244..3310
                     /label=35-ASCI-eGFP-IV-F2.6k-IF
     misc_feature    3265..3528
                     /label=35C RTR
     misc_feature    complement(3266..3528)
                     /label=35 TR(mismatch)
     misc_feature    complement(3290..3331)
                     /label=4
     primer_bind     3469..3511
                     /label=35-L1
     misc_feature    complement(3486..3527)
                     /label=1
     primer_bind     3497..3520
                     /label=Read2-35-L2
     primer_bind     3497..3520
                     /label=P7-35-L2
     misc_feature    complement(3511..3528)
                     /label=35 TR
     primer_bind     complement(3564..3581)
                     /label=M13R
                     /note="M13R"
     primer_bind     complement(3565..3581)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3607..3623)
                     /label=Rtw
                     /note="Rtw"
     terminator      3662..3693
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3694..3796
                     /label=cat promoter
                     /note="cat promoter"
     gene            3797..4657
                     /label=ampicillin_resistance_gene_ORF
                     /note="ampicillin_resistance_gene_ORF"
     CDS             4658..4849
                     /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 tcaattgtat acagtgtcgg acaaatcaat
      841 aggaccactt atcaatttat tattcactta tcatatttca tgacttaaag tgaatcaatc
      901 tttaccaaac tttcatgaaa gcactattag aatgtttact ttaagaatgt ttttaaagtt
      961 tggtgaaaag ttaagaaaca aaaaagttat gataagaaat gtaaaaataa cccaaaaatc
     1021 atgcgccaaa ataataggac cagttcagaa aattttaaaa taattatcat ttattattgt
     1081 tgctagcaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct
     1141 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     1201 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct
     1261 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     1321 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     1381 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     1441 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
     1501 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
     1561 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagccaacg ccaccatggg
     1621 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg
     1681 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg
     1741 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     1801 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     1861 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     1921 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     1981 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     2041 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     2101 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     2161 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgccatcga
     2221 tgccactaac ttctccctgt tgaaacaagc aggggatgtc gaagagaatc ccgggccaat
     2281 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2341 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2401 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2461 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     2521 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2581 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2641 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2701 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2761 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2821 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2881 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2941 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatt
     3001 aattaagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3061 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3121 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3181 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3241 ggctctatgg cttctgaggc gcgccaataa tgataaatgg ttaaaataat aaaattttct
     3301 atactggtcc tattattttg tcgcatgatt tttggggttt tttcaacatt tcttataata
     3361 acttttttgt ttgttaactt atcaacaaac tttcaaagca gactttaagc caacatccga
     3421 aaagtgcttt catgaaaatt tggtaaatat tgattcactt taggtaatga aatgtgataa
     3481 gtgaataaaa cattattaag tggtcctatt gatttgtccg acactgtata gggcccagag
     3541 acggagtcac tgccaaccga gacggtcata gctgtttcct gtgtgccgct tcctcgctca
     3601 ctgactcgct gcgctcggtc gttactgacc atttaaatca tacctgacct ccatagcaga
     3661 aagtcaaaag cctccgaccg gaggcttttg acttgatcgg cacgtaagag gttccaactt
     3721 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     3781 agctaaggaa gctaaaatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3841 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3901 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3961 tgagagttta cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4021 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4081 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctca cggatggcat
     4141 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4201 acttctggca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4261 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4321 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4381 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4441 aggatcactt ctgcgctcgg ccctcccggc tggctggttt attgctgata aatctggagc
     4501 cggtgagcgt gggtcccgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4561 catcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4621 cgctgagata ggtgcctcac tgattaagca ttggtaagtg accaaacagg aaaaaaccgc
     4681 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     4741 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     4801 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgat gagggcccaa
     4861 a
//
