LOCUS       Exported                4313 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  pCS2 with antifreeze protein terminator instead of sv40.
ACCESSION   .
VERSION     .
KEYWORDS    pCS2z
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4313)
  AUTHORS   Horstick EJ, Jordan DC, Bergeron SA, Tabor KM, Serpe M, Feldman B, 
            Burgess HA
  TITLE     Increased functional protein expression using nucleotide sequence 
            features enriched in highly expressed genes in zebrafish.
  JOURNAL   Nucleic Acids Res. 2015 Jan 27. pii: gkv035.
  PUBMED    25628360
REFERENCE   2  (bases 1 to 4313)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4313
                     /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"
     regulatory      112..121
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(609..629)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(609..627)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(648..664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(648..664)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(661..683)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    672..688
                     /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(696..726)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    741..762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(879..896)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1050..1638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1130..1149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1809..2669)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2432..2451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2670..2774)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2800..3255)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2937..2958)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3149..3168
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3329..4313
                     /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 ttcgaattca aggcctctag agccaccatg
      121 gggtaactcg agcctctagt aattaagtct cagccaccgt taactgaaca tgtcaaaacc
      181 tgtggagact gttgagattt gatgttctga aaagataaag cctataaata aaatgttgcc
      241 caaatttcct gcctgatgtt tttctttgtc tttgctacat ggctttgctg ctcggatcgg
      301 ctcactctgt gtatgccacg ttcactttgt actctccttc tcacggtagg tttattattt
      361 ttagatgtgc agttagtttc tgtgaaataa cacaccacac actgatattg tctgtgcatt
      421 gacttggtga gtgcacattg tttttgatct tgacatattt atatttgatt gatcaggtga
      481 actgtgtgaa tctaaagtgc tccatacaga tgttctgcat tgaaaatatt ctcattttat
      541 tagtggaagt gagtgtatgc cagcggccgc ggcgccaatg cattgggccc ggtacccagc
      601 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
      661 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg gagcataaag
      721 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      781 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      841 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
      901 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
      961 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1021 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1081 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1141 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1201 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1261 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1321 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1381 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1441 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     1501 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1561 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1621 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1681 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1741 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     1801 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     1861 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     1921 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     1981 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2041 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2101 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2161 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2221 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2281 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2341 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2401 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2461 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2521 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2581 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2641 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     2701 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2761 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     2821 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     2881 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     2941 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3001 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3061 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3121 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3181 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3241 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     3301 tgcgggcctc ttcgctatta cgccagtcga ccatagccaa ttcaatatgg cgtatatgga
     3361 ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga
     3421 ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc
     3481 accatgccaa ttcaatatgg cggacttggc actgtgccaa ctggggaggg gtctacttgg
     3541 cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg
     3601 catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat
     3661 ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta
     3721 tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg
     3781 cggtcccata taccatatat ggggcttcct aataccgccc atagccactc ccccattgac
     3841 gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt
     3901 atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca
     3961 ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg cccattcata
     4021 tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc
     4081 aatatctatt aatagtaact tggcaagtac attactattg gaaggacgcc agggtacatt
     4141 ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc
     4201 cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg
     4261 gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aac
//
