LOCUS       Exported                4611 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 4611)
  AUTHORS   Qinglan Biotech
  TITLE     Synthetic gene from Qinglan Biotech
  JOURNAL   Exported 04-03-2020 from Plasmap http://www.qinglanbiotech.com
REFERENCE   2  (bases 1 to 4611)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 14, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4611
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     94..113
                     /label=Ftw
                     /note="Ftw"
     primer_bind     121..138
                     /label=M13F
                     /note="M13F"
     primer_bind     122..138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    190..341
                     /label=4-5767 TR
     misc_feature    190..341
                     /label=4-5767 L-TR
     primer_bind     complement(196..233)
                     /label=Read2-4-5767-L2
     primer_bind     complement(196..233)
                     /label=P7-4-5767-L2
     misc_feature    210..259
                     /label=4
     misc_feature    210..259
                     /label=3
     primer_bind     complement(211..246)
                     /label=4-5767-L1
     misc_feature    292..341
                     /label=2
     misc_feature    292..341
                     /label=1
     primer_bind     complement(309..366)
                     /label=4-5767S-NheI-eGFP-IV-R2.6k-IF
     primer_bind     complement(309..342)
                     /label=4-5767S-ASCI-eGFP-IV-F2.6k-IF
     primer_bind     342..366
                     /label=NheI-pGK-puro/eGFP-F2.2k-IF
     primer_bind     complement(342..366)
                     /label=4-5767-NheI-eGFP-IV-R2.6k-IF
     promoter        357..856
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(627..638)
                     /label=T7ter
                     /note="T7ter"
     CDS             876..1475
                     /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             1482..1538
                     /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             1539..2258
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2239..2258
                     /label=MINI-019-SEQ-R
     polyA_signal    2286..2510
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2504..2558
                     /label=4-5767S-ASCI-eGFP-IV-F2.6k-IF
     primer_bind     2504..2525
                     /label=4-5767-ASCI-eGFP-IV-F2.6k-IF
     primer_bind     complement(2508..2525)
                     /label=ASCI-pGK-puro/eGFP-R2.2k-IF
     primer_bind     2525..2558
                     /label=4-5767S-NheI-eGFP-IV-R2.6k-IF
     misc_feature    complement(2526..2677)
                     /label=4-5767 TR
     misc_feature    2526..2677
                     /label=4-5767 R-TR
     misc_feature    complement(2526..2575)
                     /label=2
     misc_feature    complement(2526..2575)
                     /label=1
     misc_feature    complement(2608..2657)
                     /label=4
     misc_feature    complement(2608..2657)
                     /label=3
     primer_bind     2621..2656
                     /label=4-5767-L1
     primer_bind     2634..2671
                     /label=Read2-4-5767-L2
     primer_bind     2634..2671
                     /label=P7-4-5767-L2
     primer_bind     complement(2713..2730)
                     /label=M13R
                     /note="M13R"
     primer_bind     complement(2714..2730)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2756..2772)
                     /label=Rtw
                     /note="Rtw"
     rep_origin      complement(2913..3501)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3672..4532)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLRNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERKRQIAEIGAS
                     LIKHW"
     promoter        complement(join(4533..4611,1..26))
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(4574..4602)
                     /label=Amp promoter
                     /note="Amp promoter"
ORIGIN
        1 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
       61 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
      121 tgtaaaacga cggccagtcg aaccacgcaa tgcgtctcga tccgcagtgt cttgcgtctc
      181 tcaattgtac aggtgcatct caataaatta gaatgtcgtg gaaaagttca tttatttcag
      241 taattcaact caaattgtga aactcgtgta ttaaataaat tcaatgcaca cagactgaag
      301 tagtttaagt ctttggttct tttaattgtg atgattttgg cgctagcaat tctaccgggt
      361 aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac
      421 ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca
      481 accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa
      541 gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct
      601 cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg
      661 cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg
      721 gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg
      781 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc
      841 atctccgggc ctttcgacct gcagccaacg ccaccatggg gaccgagtac aagcccacgg
      901 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg
      961 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg
     1021 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     1081 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     1141 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     1201 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     1261 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     1321 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     1381 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     1441 cgcgcacctg gtgcatgacc cgcaagcccg gtgccatcga tgccactaac ttctccctgt
     1501 tgaaacaagc aggggatgtc gaagagaatc ccgggccaat ggtgagcaag ggcgaggagc
     1561 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1621 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1681 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1741 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1801 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1861 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1921 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1981 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2041 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2101 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     2161 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2221 ccgggatcac tctcggcatg gacgagctgt acaagtaatt aattaagctc gctgatcagc
     2281 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2341 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2401 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2461 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2521 gcgccgccaa aatcatcaca attaaaagaa ccaaagactt aaactacttc agtctgtgtg
     2581 cattgaattt atttaataca cgagtttcac aatttgagtt gaattactga aataaatgaa
     2641 cttttccacg acattctaat ttattgagat gcacctgtag ggcccagaga cggagtcact
     2701 gccaaccgag acggtcatag ctgtttcctg tgtgccgctt cctcgctcac tgactcgctg
     2761 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2821 cccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2881 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2941 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3001 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3061 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3121 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3181 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3241 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3301 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3361 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3421 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3481 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3541 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3601 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3661 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctcttt
     3721 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3781 ccatctggcc ccagtgctgc aataataccg cgggacccac gctcaccggc tccagattta
     3841 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3901 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3961 agtttgcgca acgttgttgc catcgctaca ggcatcgtgg tatcacgctc gtcgtttggt
     4021 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4081 cgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgcc
     4141 gtgttatcac tcatggttat ggcagcacta cataattctc ttactgtcat gccatccgta
     4201 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4261 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4321 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4381 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4441 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4501 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4561 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta g
//
