LOCUS       Exported                5630 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  in vitro transcription.
ACCESSION   .
VERSION     .
KEYWORDS    pCS2+ EB3-mKate2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5630)
  AUTHORS   Strzyz PJ, Lee HO, Sidhaye J, Weber IP, Leung LC, Norden C
  TITLE     Interkinetic nuclear migration is centrosome independent and ensures
            apical cell division to maintain tissue integrity.
  JOURNAL   Dev Cell. 2015 Jan 26;32(2):203-19. doi: 
            10.1016/j.devcel.2014.12.001. Epub 2015 Jan 15.
  PUBMED    25600237
REFERENCE   2  (bases 1 to 5630)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5630
                     /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             946..1644
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     promoter        complement(1660..1677)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1682..1816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1706..1725)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1760..1779
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1929..1949)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1929..1947)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1968..1984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1968..1984)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1981..2003)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1992..2008
                     /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(2016..2046)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2061..2082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2199..2216)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2370..2958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2450..2469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3129..3989)
                     /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     3752..3771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3990..4094)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4120..4575)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4257..4278)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4469..4488
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4653..5630
                     /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 tttttgcaat cgatatggcc gtcaatgtgt actccacatc tgtgaccagt
      121 gaaaatctga gtcgccatga tatgcttgca tgggtcaacg actccctgca cctcaactat
      181 accaagatag aacagctttg ttcaggggca gcctactgcc agttcatgga catgctcttc
      241 cccggctgtg tgcacttgag gaaagtgaag ttccaggcca aactagagca tgaatacatc
      301 cacaacttca aggtgctgca agcagctttc aagaagatgg gtgttgacaa aatcattcct
      361 gtagagaaat tagtgaaagg aaaattccaa gataattttg agtttattca gtggtttaag
      421 aaattctttg acgcaaacta tgatggaaag gattacaacc ctctgctggc gcggcagggc
      481 caggacgtag cgccacctcc taacccaggt gatcagatct tcaacaaatc caagaaactc
      541 attggcacag cagttccaca gaggacgtcc cccacaggcc caaaaaacat gcagacctct
      601 ggccggctga gcaatgtggc ccccccctgc attctccgga agaatcctcc atcagcccga
      661 aatggcggcc atgagactga tgcccaaatt cttgaactca accaacagct ggtggacttg
      721 aagctgacag tggatgggct ggagaaggaa cgtgacttct acttcagcaa acttcgtgac
      781 atcgagctca tctgccagga gcatgaaagt gaaaacagcc ctgttatctc aggcatcatt
      841 ggcatcctct atgccacaga ggaaggattc gcaccccctg aggacgatga gattgaagag
      901 catcaacaag aagaccagga cgagtacacc ggttcatcgg gatccatggt gagcgagctg
      961 attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc
     1021 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
     1081 gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac
     1141 ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc
     1201 cccgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct
     1261 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
     1321 aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggcctccacc
     1381 gagaccctgt accccgctga cggcggcctg gaaggcagag ccgacatggc cctgaagctc
     1441 gtgggcgggg gccacctgat ctgcaacttg aagaccacat acagatccaa gaaacccgct
     1501 aagaacctca agatgcccgg cgtctactat gtggacagaa gactggaaag aatcaaggag
     1561 gccgacaaag agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc
     1621 cctagcaaac tggggcacag atgactcgag cctctagaac tatagtgagt cgtattacgt
     1681 agatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1741 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1801 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1861 ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc ggtacccagc
     1921 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     1981 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg gagcataaag
     2041 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2101 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2161 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2221 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2281 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2341 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2401 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2461 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2521 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2581 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2641 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2701 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2761 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2821 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2881 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2941 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3001 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3061 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3121 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3181 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3241 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3301 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3361 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3421 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3481 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3541 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3601 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3661 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3721 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3781 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3841 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3901 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3961 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4021 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4081 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     4141 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4201 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4261 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4321 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4381 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4441 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     4501 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4561 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4621 tgcgggcctc ttcgctatta cgccagtcga ccgccaattc aatatggcgt atatggactc
     4681 atgccaattc aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc
     4741 gtgccaattc aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc
     4801 atgccaattc aatatggcgg acctggcact gtgccaactg gggaggggtc tacttggcac
     4861 ggtgccaagt ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat
     4921 ggtgccaata atggcggcca tattggctat atgccaggat caatatatag gcaatatcca
     4981 atatggccct atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc
     5041 catatagtat tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg
     5101 tcccatatac catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc
     5161 aatggtctct atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata
     5221 tggcgcctcc cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg
     5281 acgtcaatag gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc
     5341 gttctcacgc cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat
     5401 atctattaat agtaacttgg caagtacatt actattggaa gtacgccagg gtacattggc
     5461 agtactccca ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc
     5521 attgacgtca atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg
     5581 gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg tttagggaac
//
