LOCUS       Exported                4924 bp ds-DNA     circular SYN 16-OCT-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4924)
  AUTHORS   Qinglan Biotech
  TITLE     Synthetic gene from Qinglan Biotech
  JOURNAL   Exported 15-11-2019 from Plasmap http://www.qinglanbiotech.com
REFERENCE   2  (bases 1 to 4924)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 23, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4924
                     /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"
     rep_origin      complement(51..638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     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    818..819
                     /label=TSD
     misc_feature    820..1137
                     /label=59LTR
     misc_feature    820..848
                     /label=predictive TIR
     promoter        1159..1658
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1429..1440)
                     /label=T7ter
                     /note="T7ter"
     CDS             1678..2277
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     gene            1683..2277
                     /label=puromycin_transacetylase
                     /note="puromycin_transacetylase"
     CDS             2284..2340
                     /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"
     gene            2341..3060
                     /label=eGFP
                     /note="eGFP"
     CDS             2341..3060
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3088..3312
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3320..3327
                     /locus_tag="MCS(5)"
                     /label=MCS(5)
     misc_feature    3334..3591
                     /label=59RTR
     misc_feature    complement(3563..3591)
                     /label=predictive TIR
     misc_feature    3592..3593
                     /label=TSD
     primer_bind     complement(3627..3644)
                     /label=M13R
                     /note="M13R"
     primer_bind     complement(3628..3644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3670..3686)
                     /label=Rtw
                     /note="Rtw"
     terminator      3725..3756
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3757..3859
                     /label=cat promoter
                     /note="cat promoter"
     gene            3860..4720
                     /label=ampicillin_resistance_gene_ORF
                     /note="ampicillin_resistance_gene_ORF"
     CDS             3860..4720
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             4721..4912
                     /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 actgctcaaa aaaattagag
      841 gaacactttt taatcagagt atagcaacct atcagtcaaa cttctgggat attgatctgg
      901 tcagttaagt agcagagggg gttgttaatc agtttctgct gctttgttgt taatgaaatc
      961 aacaacagga gcatcagagg ggcaacaatg agacggcccc caaaacagga atggctttcc
     1021 aggttgaggc cactgatact tttttccctc ctcatctctt ttggctgatt ttttactgac
     1081 tgactttcta ctgctgtagt tattcatttg gcttggatca acatctctgt tgatagcacc
     1141 ggtgctagca attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
     1201 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
     1261 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
     1321 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
     1381 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
     1441 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
     1501 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
     1561 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
     1621 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccaa cgccaccatg
     1681 gggaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta
     1741 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac
     1801 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     1861 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     1921 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     1981 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     2041 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     2101 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     2161 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     2221 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgccatc
     2281 gatgccacta acttctccct gttgaaacaa gcaggggatg tcgaagagaa tcccgggcca
     2341 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2401 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2461 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2521 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     2581 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2641 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2701 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2761 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     2821 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2881 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2941 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3001 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     3061 ttaattaagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3121 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3181 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3241 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3301 tgggctctat ggcttctgag gcgcgccctc gaggaatcat tttgagttgc tacaatgaca
     3361 ttttggcaaa atggaccagt ctgctgcatc attttttcac tttcattttt ggggtgtctt
     3421 tgatttcccc cctctatagg gtgatcattt tcatttctat caaattatgt ggcatcattt
     3481 tgttactaat acatcaccta ctttttatca ggaaagatat tcaagatcat tttccccccc
     3541 gtttagatct gatgtgtttt cgaagtgttc ctctaatttt tttgagcagt gtagggccca
     3601 gagacggagt cactgccaac cgagacggtc atagctgttt cctgtgtgcc gcttcctcgc
     3661 tcactgactc gctgcgctcg gtcgttactg accatttaaa tcatacctga cctccatagc
     3721 agaaagtcaa aagcctccga ccggaggctt ttgacttgat cggcacgtaa gaggttccaa
     3781 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
     3841 aggagctaag gaagctaaaa tgagtattca acatttccgt gtcgccctta ttcccttttt
     3901 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3961 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4021 ccttgagagt ttacgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4081 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4141 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc tcacggatgg
     4201 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4261 cttacttctg gcaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4321 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4381 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4441 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4501 tgcaggatca cttctgcgct cggccctccc ggctggctgg tttattgctg ataaatctgg
     4561 agccggtgag cgtgggtccc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4621 ccgcatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4681 gatcgctgag ataggtgcct cactgattaa gcattggtaa gtgaccaaac aggaaaaaac
     4741 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
     4801 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
     4861 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gatgagggcc
     4921 caaa
//
