LOCUS       Exported                4897 bp ds-DNA     circular SYN 19-AUG-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4897)
  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 4897)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 14, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4897
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1184..3357
                     /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..1183
                     /label=14-LTR
     misc_feature    820..1019
                     /label=14 TR
     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(988..1019)
                     /label=14-ASCI-copGFP-IV-F2.6k-IF
     primer_bind     complement(990..1020)
                     /label=14-SpeI-copGFP-IV-R2.6k-IF
     misc_feature    1104..1153
                     /label=5
     misc_feature    1184..1189
                     /locus_tag="MCS"
                     /label=MCS
     primer_bind     1191..1215
                     /label=XbaI-CMV-F1.8K(W)
     enhancer        1196..1215
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(1215..1230)
                     /label=Primer 1
     promoter        1216..2394
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     complement(1331..1354)
                     /label=EF1a-jun-R
     intron          1447..2385
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(2382..2422)
                     /label=pmax-KpnI-EF1A-R1.2K-IF
     misc_feature    2402..2407
                     /label=MCS
     primer_bind     complement(2423..2438)
                     /label=2-NcoI-EF1a-R-IF
     primer_bind     complement(2423..2438)
                     /label=2-NcoI/ClaI-EF1a-R-IF
     CDS             2436..3101
                     /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(2529..2550)
                     /label=CopGFP-seq-R
     misc_feature    3109..3114
                     /label=MCS(1)
     misc_feature    3117..3317
                     /label=SV40 polyA
     primer_bind     3325..3343
                     /label=IV-ASCI-CopGFP-F
     primer_bind     3327..3396
                     /label=14-ASCI-copGFP-IV-F2.6k-IF
     misc_feature    3352..3357
                     /locus_tag="MCS(4)"
                     /label=MCS(4)
     misc_feature    3358..3564
                     /label=14-RTR
     primer_bind     3362..3394
                     /label=14-SpeI-copGFP-IV-R2.6k-IF
     misc_feature    complement(3365..3564)
                     /label=14 TR
     misc_feature    complement(3382..3431)
                     /label=4
     misc_feature    complement(3382..3431)
                     /label=3
     misc_feature    complement(3484..3533)
                     /label=2
     misc_feature    complement(3484..3533)
                     /label=1
     primer_bind     complement(3600..3617)
                     /label=M13R
                     /note="M13R"
     primer_bind     complement(3643..3659)
                     /label=Rtw
                     /note="Rtw"
     terminator      3698..3729
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3730..3832
                     /label=cat promoter
                     /note="cat promoter"
     gene            3833..4693
                     /label=ampicillin_resistance_gene_ORF
                     /note="ampicillin_resistance_gene_ORF"
     CDS             4694..4885
                     /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 attgaacaca
     1021 tgaagataac aaggtgcaaa atggcataga aagtcaggag atctgtcagt attgagagaa
     1081 aaaccctgct ccctatcagt actaattgat atcagctgct ttagtcctaa ttgatggcct
     1141 ataaaggctt ctcattactc aggaggcaca caggaaagac ttcaccggtg ctagaggcat
     1201 tgattattga ctagtggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     1261 cccgagaagt tggggggagg ggtcggcaat tgacccggtg cctagagaag gtggcgcggg
     1321 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     1381 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     1441 acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct
     1501 tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg
     1561 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
     1621 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
     1681 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
     1741 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt
     1801 tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg
     1861 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctcgccgg
     1921 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
     1981 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg
     2041 agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
     2101 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccactgagta ccgggcgccg
     2161 tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
     2221 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     2281 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     2341 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgagggatc
     2401 cggtaccgaa gccgctagcg ctaccggtcg ccaccatgcc cgccatgaag atcgagtgcc
     2461 gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga gagggcaccc
     2521 ccgagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg accttcagcc
     2581 cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc taccccagcg
     2641 gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac acccgcatcg
     2701 agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac gaggccggcc
     2761 gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc gtgatcttca
     2821 ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg ggcgataacg
     2881 tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac tacagcttcg
     2941 tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg cagaacgggg
     3001 gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag ctgggcatcg
     3061 tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatctcga gctcgatgag
     3121 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3181 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3241 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     3301 ctctacaaat gtggtactta agagggggag accaaagggc gagggcgcgc cctcgaggta
     3361 aagtgtgttc aatacttatt ccctgtgtca tttcacttta tttattatga ctcaacttgt
     3421 atacttaaat gttctgattt ctttgtatga attcaatatt tggcttgatg gctacatctg
     3481 gtaggaattt tgtgtcagta gcccacttag aaatcccctt actgataaaa atgctgatgt
     3541 gtcaaatact tattttcccc gctgtagggc ccagagacgg agtcactgcc aaccgagacg
     3601 gtcatagctg tttcctgtgt gccgcttcct cgctcactga ctcgctgcgc tcggtcgtta
     3661 ctgaccattt aaatcatacc tgacctccat agcagaaagt caaaagcctc cgaccggagg
     3721 cttttgactt gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact
     3781 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatgagtat
     3841 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3901 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3961 ttacatcgaa ctggatctca acagcggtaa gatccttgag agtttacgcc ccgaagaacg
     4021 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4081 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4141 ctcaccagtc acagaaaagc atctcacgga tggcatgaca gtaagagaat tatgcagtgc
     4201 tgccataacc atgagtgata acactgcggc caacttactt ctggcaacga tcggaggacc
     4261 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4321 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4381 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4441 acaattaata gactggatgg aggcggataa agttgcagga tcacttctgc gctcggccct
     4501 cccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt cccgcggtat
     4561 cattgcagca ctggggccag atggtaagcc ctcccgcatc gtagttatct acacgacggg
     4621 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4681 taagcattgg taagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag
     4741 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga
     4801 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt
     4861 cggtgatgac ggtgaaaacc tctgatgagg gcccaaa
//
