LOCUS       ER_CFAST_10_in_p        5738 bp DNA     circular SYN 29-SEP-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5738)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Oct 11, 2023 from SnapGene Viewer 7.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5738
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    895..929
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     928..1025
                     /label=Primer 1
     primer_bind     928..967
                     /label=Primer 1 (1)
     regulatory      935..944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             944..973
                     /codon_start=1
                     /label=CFAST10
                     /translation="GDSYWVFVKR"
     CDS             1022..1228
                     /codon_start=1
                     /label=ER-targeting sequence
                     /translation="RVFLALPIIMVVAFSMCIICLLMAGDTWTETLAYVLFWGVASIGT
                     FFIILYNGKDFVDAPRLVQKEKID"
     primer_bind     complement(1177..1228)
                     /label=Primer 2
     primer_bind     complement(1213..1267)
                     /label=Primer 2 (1)
     CDS             1235..1258
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1290..1312
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(1332..1349)
                     /label=bGH reverse primer
     polyA_signal    1338..1562
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1608..2036
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2050..2379
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2230..2365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2446..3240
                     /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)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3414..3535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3986..4571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4742..5602)
                     /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"
     promoter        complement(5603..5707)
                     /gene="bla"
                     /label=AmpR promoter
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 aagcttggta ccgagctcgg atccgccgcc atgggtgaca gctattgggt
      961 ctttgtgaaa cgggaattcg gcggagatgg cggatctggc ggcggagata tcagcaagct
     1021 gagagtgttc ctggccctgc ccatcatcat ggtggtggcc ttttctatgt gcatcatctg
     1081 tctgctgatg gcaggcgaca cctggacaga gaccctggcc tacgtgctgt tttggggcgt
     1141 ggccagcatc ggcacattct ttatcatcct gtataacggc aaggacttcg tggacgcccc
     1201 cagactggtg cagaaagaga agatcgacga tatcgattac aaggatgacg acgataagtg
     1261 atctagaccg tcgacacgga attcattgcg gccgctcgag tctagagggc ccgtttaaac
     1321 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1381 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1441 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1501 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1561 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1621 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1681 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1741 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1801 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1861 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1921 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1981 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2041 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2101 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2161 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2221 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2281 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2341 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2401 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2461 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2521 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2581 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2641 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2701 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2761 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     2821 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     2881 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     2941 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3001 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3061 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3121 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3181 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3241 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3301 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3361 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     3421 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3481 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3541 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3601 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3661 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3721 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3781 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3841 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3901 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3961 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4021 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4081 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4141 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4201 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4261 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4321 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4381 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4441 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4501 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4561 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4621 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4681 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4741 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4801 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4861 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4921 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4981 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5041 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5101 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5161 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5221 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5281 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5341 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5401 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5461 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5521 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5581 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5641 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5701 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
