LOCUS       eIscB        7129 bp DNA     circular SYN 17-SEP-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    eIscB
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7129)
  AUTHORS   GenScript
  TITLE     Direct Submission
  JOURNAL   Exported Sep 17, 2024 from SnapGene 7.0.2
            https://www.snapgene.com
COMMENT     http://www.genscript.com/
            ORIGDB|GenBank
FEATURES             Location/Qualifiers
     source          1..7129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             932..952
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             2450..2470
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             2477..2524
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /note="color: #993366"
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2531..2611
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     polyA_signal    2726..2950
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      2996..3424
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3438..3767
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3618..3753
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             3834..4628
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4802..4923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(4972..4988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    4996..5012
                     /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)."
                     /note="color: #31849b"
     promoter        complement(5020..5050)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5020 .. 5026 / #ffffff / -10
                        2: 5027 .. 5044 / #ffffff
                        3: 5045 .. 5050 / #ffffff / -35"
     protein_bind    5065..5086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(5374..5962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6133..6993)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6133 .. 6924 / #ccffcc
                        2: 6925 .. 6993 / #ccffcc / signal sequence
                     Cleavage site after base 6924"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6994..7098)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgggatc tccaaaaaag aaacggaagg tgggatctat
      961 ggccgtggtc tacgtgatct ctaagtccgg caaaccactg atgcccacca ccagatgcgg
     1021 ccacgtgcgg atcttgctca aggaaggcaa ggctagagtg gtggaaagaa agcccttcac
     1081 catccagctg acctacgagt ctgccgagga aacccagcct ctggtgctgg gcattgaccc
     1141 tggcagaacc aatatcggta tgagcgtggt gaccgagagc ggagagagcg tgttcaacgc
     1201 ccaaatcaga acaagaaaca aggacgtgcc taagctgatg aagagaagaa agcagtatag
     1261 gatggcccac agaaggctga agcggcggtg caaaagacgg cggagagcca aggctgccgg
     1321 caccgccttc gaggagggcg aaaagcaaag actgctgcct ggctgcttca agcctattac
     1381 atgcaagagc atccggaaca aggaagccag attcaacaac cgcaagcggc ctagaggatg
     1441 gctgacacct accgccaatc acctgctggt cacccacctg aacgtggtta agaaggtgca
     1501 gaagatcctg cccgtggcca aggtggtgct ggagctgaat cggttcagct ttatggccat
     1561 gaacaacccc aaggttcaga gatggcagta ccagcggggc cctctgtacg gcaaaggcag
     1621 cgtggaagag gccgtgtcca tgcagcagga tggacactgc ctgttttgta aacacggcat
     1681 cgaccactac caccacgtgg tccccagaag aaagaacggc agcgagacac tggaaaacag
     1741 agtgggcctt tgtgaagaac accatagact ggtgcacaca gataaggagt gggaggccaa
     1801 cctggccagc aagaagtctg gaatgaacaa gaaataccac gccctgtctg tgctgaacca
     1861 gatcatcccc tacctggctg atcagctggc cgatatgttc cccggaaatt tctgcgtgac
     1921 aagcggccaa gacacctatc tgtttcggga agagcatggc atccctaagg accactacct
     1981 ggacgcctac tgcatcgcct gcagcgcact gaccgacgcc aagaaggtga gcagccctaa
     2041 gggcagacca tacatggtgc accagttcag aaggcatgat agacaggcct gtcacaaggc
     2101 caatctgaac cggagctact acatgggcgg caagctcgtt gccaccaacc ggcacaaagc
     2161 tatggaccag aaaactgaca gcctggaaga gtacagagcc gctcacagcg ccgctgacgt
     2221 gtctaaactg accgtgaagc acccttctgc tcagtacaag gatatgagca gaatcatgcc
     2281 tggcagcatc ctggtgagcg gcgagggcaa actgttcaca ctgagcagat ctgagggaag
     2341 aaacaaaggc caggtgaact acttcgtgtc caccgagggc atcaagtact gggccagaaa
     2401 gtgccagtat ctgcggaaca acggcggact gcagatctac gtgggatccc ctaagaaaaa
     2461 acgaaaagtt ggatccaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa
     2521 aaagggatct tacccatacg atgttccaga ttacgcttac ccatacgatg ttccagatta
     2581 cgcttaccca tacgatgttc cagattacgc ttgaggtacc gagctcggat ccactagtcc
     2641 agtgtggtgg aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc
     2701 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2761 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2821 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2881 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2941 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3001 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3061 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3121 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3181 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3241 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3301 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3361 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3421 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3541 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3601 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3661 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3721 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3781 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3841 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3901 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3961 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4021 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4081 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4141 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4201 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4261 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4321 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4381 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4441 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4501 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4561 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4621 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4681 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4741 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4801 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4861 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4921 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4981 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5041 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5101 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5161 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5221 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5281 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5341 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5401 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5461 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5521 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5581 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5641 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5701 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5761 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5821 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5881 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5941 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6001 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6061 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6121 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6181 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6241 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6301 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6361 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6421 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6481 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6541 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6601 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6661 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6721 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6781 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6841 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6901 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6961 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7021 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7081 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
