LOCUS       Exported                5504 bp ds-DNA     circular SYN 06-JAN-2022
DEFINITION  enables eukaryotic expression of N-terminally FLAG-HA-tagged fusion 
            proteins.
ACCESSION   .
VERSION     .
KEYWORDS    FLAG-HA-pcDNA3.1-
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  AUTHORS   Horn M, Geisen C, Cermak L, Becker B, Nakamura S, Klein C, Pagano M,
            Antebi A
  TITLE     DRE-1/FBXO11-Dependent Degradation of BLMP-1/BLIMP-1 Governs C. 
            elegans Developmental Timing and Maturation.
  JOURNAL   Dev Cell. 2014 Mar 4. pii: S1534-5807(14)00069-0. doi: 
            10.1016/j.devcel.2014.01.028.
  PUBMED    24613396
REFERENCE   2  (bases 1 to 5504)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 6, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5504
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      169..304
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     231..250
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             385..1179
                     /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))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(439..458)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1049..1068
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1353..1474
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1390..1409)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1444..1463
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1523..1539)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1523..1539)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1536..1558)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1547..1563
                     /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(1571..1601)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1616..1637
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1754..1771)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1925..2513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2005..2024)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2684..3544)
                     /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     3307..3326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3545..3649)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3724..3743)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3915..4294
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4295..4498
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4449..4469
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4543..4562
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4543..4561
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4579..4588
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4588..4611
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4627..4653
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4775..4792)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4781..5005
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5051..5479
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5138..5157)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5348..5369
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5488..4)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5493..318
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
       61 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
      121 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
      181 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
      241 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
      301 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
      361 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
      421 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
      481 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
      541 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
      601 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
      661 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
      721 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
      781 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
      841 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
      901 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
      961 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     1021 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     1081 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     1141 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     1201 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     1261 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     1321 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     1381 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1441 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     1501 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1561 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1621 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1681 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1741 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1801 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1861 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1921 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1981 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2041 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2101 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2161 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2221 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2281 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2341 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2401 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2461 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2521 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2581 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2641 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2701 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2761 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2821 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2881 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     2941 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3001 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3061 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3121 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3181 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3241 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3301 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3361 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3421 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3481 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3541 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3601 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3661 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     3721 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     3781 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     3841 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     3901 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     3961 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     4021 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     4081 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     4141 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     4201 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     4261 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     4321 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     4381 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     4441 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
     4501 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
     4561 ggagacccaa gctggctagc caccatggac tacaaagacg atgacgataa agggggcggt
     4621 ggaggttacc catacgatgt tccagattac gctggtggag gtggaggttc tagactcgag
     4681 cggccgccac tgtgctggat atctgcagaa ttccaccaca ctggactagt ggatccgagc
     4741 tcggtaccaa gcttaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag
     4801 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     4861 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     4921 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     4981 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     5041 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5101 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     5161 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5221 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5281 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5341 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5401 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5461 caaaaattta acgcgaatta attctgtgga atgtgtgtca gtta
//
