LOCUS       Exported                5324 bp ds-DNA     circular SYN 22-NOV-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5324)
  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 5324)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 14, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5324
                     /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..1442
                     /label=35C LTR
     misc_feature    820..1081
                     /label=35 TR(mismatch)
     misc_feature    820..837
                     /label=35 TR
     misc_feature    820..837
                     /label=35 TR
     misc_feature    821..862
                     /label=3
     misc_feature    1016..1057
                     /label=2
     primer_bind     complement(1037..1082)
                     /label=35-SpeI-copGFP-IV-R2.6k-IF
     primer_bind     1419..1442
                     /label=35-LTIR-F
     misc_feature    1443..1448
                     /locus_tag="MCS"
                     /label=MCS
     primer_bind     1450..1474
                     /label=XbaI-CMV-F1.8K(W)
     enhancer        1455..1474
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1475..2653
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     complement(1590..1613)
                     /label=EF1a-jun-R
     intron          1706..2644
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(2641..2681)
                     /label=pmax-KpnI-EF1A-R1.2K-IF
     misc_feature    2661..2666
                     /label=MCS
     primer_bind     complement(2682..2697)
                     /label=2-NcoI/ClaI-EF1a-R-IF
     primer_bind     complement(2682..2697)
                     /label=2-NcoI-EF1a-R-IF
     CDS             2695..3360
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     primer_bind     complement(2788..2809)
                     /label=CopGFP-seq-R
     misc_feature    3368..3373
                     /label=MCS(1)
     misc_feature    3376..3576
                     /label=SV40 polyA
     primer_bind     complement(3585..3610)
                     /label=ASCI-CMV-R1.8K(W)
     misc_feature    3611..3616
                     /locus_tag="MCS(4)"
                     /label=MCS(4)
     misc_feature    3617..3991
                     /label=35C RTR
     primer_bind     complement(3619..3638)
                     /label=35-RTIR-R
     primer_bind     3728..3773
                     /label=35-ASCI-CopGFP-IV-F2.6k-IF
     misc_feature    complement(3729..3991)
                     /label=35 TR(mismatch)
     misc_feature    complement(3753..3794)
                     /label=4
     misc_feature    complement(3949..3990)
                     /label=1
     misc_feature    complement(3974..3991)
                     /label=35 TR
     misc_feature    complement(3974..3991)
                     /label=35 TR
     primer_bind     complement(4027..4044)
                     /label=M13R
                     /note="M13R"
     primer_bind     complement(4028..4044)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4070..4086)
                     /label=Rtw
                     /note="Rtw"
     terminator      4125..4156
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        4157..4259
                     /label=cat promoter
                     /note="cat promoter"
     gene            4260..5120
                     /label=ampicillin_resistance_gene_ORF
                     /note="ampicillin_resistance_gene_ORF"
     CDS             5121..5312
                     /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 taaaacacag tttttgatgt tattgagttg tcggataact taacttaaat tagtgaacta
     1141 atgcgtattc attttttttg tagaaacgct tagcgcagtg gcaattcgct gtgtctttaa
     1201 ttaacatgct ctaatttact aatgcaagaa tattacggca ctcgcttaag ggttttgagc
     1261 tccctttgca ctgagagacg aaatagtggt gagtaccttc gtctatgggt ataaaagcgc
     1321 gtcacagcgc tggatcgctt acactgtgtt ctaagccgtt gcctagaaca cttcacacga
     1381 tccattagct cgtgtatttt tgggtgcact acttcaagcg cttgattgaa ggaattccaa
     1441 cgaccggtgc tagaggcatt gattattgac tagtggctcc ggtgcccgtc agtgggcaga
     1501 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gacccggtgc
     1561 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     1621 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     1681 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
     1741 ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat
     1801 tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg
     1861 agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg
     1921 aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa
     1981 tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca
     2041 agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc
     2101 ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg
     2161 gtagtctcaa gctcgccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc
     2221 gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct
     2281 tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg
     2341 tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc
     2401 cactgagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc
     2461 gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga
     2521 gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga
     2581 gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt
     2641 tcaggtgtcg tgagggatcc ggtaccgaag ccgctagcgc taccggtcgc caccatgccc
     2701 gccatgaaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg
     2761 ggcggcggag agggcacccc cgagcagggc cgcatgacca acaagatgaa gagcaccaaa
     2821 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac
     2881 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc
     2941 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc
     3001 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc
     3061 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg
     3121 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac
     3181 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc
     3241 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc
     3301 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc
     3361 agatctcgag ctcgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3421 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3481 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3541 tttttaaagc aagtaaaacc tctacaaatg tggtacttaa gagggggaga ccaaagggcg
     3601 agggcgcgcc ctcgagatgc taccaagtta gcccaggata aggctgtttt gttattttct
     3661 ttagtttttc aaatgtacgg gctggccaca ttgcgaatta aaagccggag ctgtaattat
     3721 ttataggcaa taatgataaa tggttaaaat aataaaattt tctatactgg tcctattatt
     3781 ttgtcgcatg atttttgggg ttttttcaac atttcttata ataacttttt tgtttgttaa
     3841 cttatcaaca aactttcaaa gcagacttta agccaacatc cgaaaagtgc tttcatgaaa
     3901 atttggtaaa tattgattca ctttaggtaa tgaaatgtga taagtgaata aaacattatt
     3961 aagtggtcct attgatttgt ccgacactgt atagggccca gagacggagt cactgccaac
     4021 cgagacggtc atagctgttt cctgtgtgcc gcttcctcgc tcactgactc gctgcgctcg
     4081 gtcgttactg accatttaaa tcatacctga cctccatagc agaaagtcaa aagcctccga
     4141 ccggaggctt ttgacttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa
     4201 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
     4261 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4321 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4381 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt ttacgccccg
     4441 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4501 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4561 ttgagtactc accagtcaca gaaaagcatc tcacggatgg catgacagta agagaattat
     4621 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg gcaacgatcg
     4681 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4741 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4801 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4861 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggatca cttctgcgct
     4921 cggccctccc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtccc
     4981 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgcatcgta gttatctaca
     5041 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5101 cactgattaa gcattggtaa gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
     5161 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
     5221 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg
     5281 cgcgtttcgg tgatgacggt gaaaacctct gatgagggcc caaa
//
