LOCUS       pCS2FLAG                4151 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCS2FLAG
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4151)
  TITLE     Klein lab unpublished plasmids
REFERENCE   2  (bases 1 to 4151)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 10, 2018 from SnapGene Server 1.1.58
            //www.snapgene.com
COMMENT     This file is created by Vector NTI
            //gt12365.cn/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|gt12365.cn|
COMMENT     VNTNAME|pCS2FLAG|
FEATURES             Location/Qualifiers
     source          1..4151
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             103..150
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     promoter        complement(178..195)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    200..334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(224..243)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     278..297
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(447..467)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(447..465)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(486..502)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(486..502)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(499..521)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    510..526
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(534..564)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    579..600
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(717..734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(888..1476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(968..987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1647..2507)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     primer_bind     2270..2289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2508..2612)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2638..3093)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2775..2796)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2987..3006
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3167..4151
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tttaaagcta tggattacaa ggatgatgat
      121 gataaggatt acaaggatga tgatgataag aattcaaggc ctctcgagcc tctagaacta
      181 tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt ttggacaaac
      241 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
      301 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
      361 gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca
      421 ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
      481 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
      541 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
      601 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
      661 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
      721 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
      781 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      841 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      901 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      961 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1021 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1081 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1141 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1201 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1261 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1321 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1381 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1441 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1501 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1561 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1621 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1681 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1741 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1801 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1861 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1921 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1981 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2041 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2101 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2161 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2221 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2281 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2341 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2401 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2461 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2521 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2581 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     2641 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     2701 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2761 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2821 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     2881 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2941 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3001 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3061 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     3121 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc atagccaatt
     3181 caatatggcg tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt
     3241 caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt
     3301 caatatggcg gacttggcac catgccaatt caatatggcg gacttggcac tgtgccaact
     3361 ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag
     3421 tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga
     3481 tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa
     3541 tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag
     3601 atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat
     3661 agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat
     3721 gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc
     3781 cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg
     3841 gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg
     3901 cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga
     3961 aggacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg
     4021 atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg
     4081 ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc
     4141 gtttagggaa c
//