LOCUS       sequence-169437-        6074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6074)
  AUTHORS   Okumura F, Zou W, Zhang DE
  TITLE     ISG15 modification of the eIF4E cognate 4EHP enhances cap 
            structure-binding activity of 4EHP.
  JOURNAL   Genes Dev. 2007 Feb 1. 21(3):255-60.
  PUBMED    17289916
REFERENCE   2  (bases 1 to 6074)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genes Dev. 
            2007 Feb 1. 21(3):255-60."
COMMENT     Sequence Label: pHA-eIF4E
FEATURES             Location/Qualifiers
     source          1..6074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..30
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(727..745)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(728..745)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(765..782)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    771..995
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1041..1469
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1128..1147)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1338..1359
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1478..1498)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1483..1812
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1663..1798
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1725..1744
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1879..2673
                     /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(1933..1952)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2543..2562
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2847..2968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2884..2903)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2938..2957
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3017..3033)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3017..3033)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3030..3052)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3041..3057
                     /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(join(3065..3071,3072..3089,3090..3095))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3110..3131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3248..3265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3419..4007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3499..3518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4178..4969,4970..5038))
                     /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     4801..4820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5039..5143)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5218..5237)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5409..5788
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5789..5992
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5943..5963
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6037..6056
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6037..6055
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atgtacccat acgacgtccc agactacgct ggaagcttgg taccgatggc gactgtcgaa
       61 ccggaaacca cccctactcc taatcccccg actacagaag aggagaaaac ggaatctaat
      121 caggaggttg ctaacccaga acactatatt aaacatcccc tacagaacag atgggcactc
      181 tggtttttta aaaatgataa aagcaaaact tggcaagcaa acctgcggct gatctccaag
      241 tttgatactg ttgaagactt ttgggctctg tacaaccata tccagttgtc tagtaattta
      301 atgcctggct gtgactactc actttttaag gatggtattg agcctatgtg ggaagatgag
      361 aaaaacaaac ggggaggacg atggctaatt acattgaaca aacagcagag acgaagtgac
      421 ctcgatcgct tttggctaga gacacttctg tgccttattg gagaatcttt tgatgactac
      481 agtgatgatg tatgtggcgc tgttgttaat gttagagcta aaggtgataa gatagcaata
      541 tggactactg aatgtgaaaa cagagaagct gttacacata tagggagggt atacaaggaa
      601 aggttaggac ttcctccaaa gatagtgatt ggttatcagt cccacgcaga cacagctact
      661 aagagcggct ccaccactaa aaataggttt gttgtttaac tcgagcatgc atctagaggg
      721 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
      781 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
      841 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
      901 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
      961 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     1021 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1081 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1141 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1201 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1261 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1321 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1381 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1441 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     1501 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1561 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     1621 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     1681 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     1741 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     1801 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     1861 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     1921 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     1981 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2041 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     2101 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     2161 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     2221 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     2281 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     2341 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     2401 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     2461 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     2521 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     2581 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     2641 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     2701 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     2761 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     2821 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     2881 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2941 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     3001 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3061 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3121 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3181 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3241 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3301 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3361 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3421 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3481 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3541 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3601 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3661 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3721 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3781 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3841 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3901 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3961 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4021 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4081 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4141 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4201 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4261 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4321 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4381 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4441 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4501 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4561 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4621 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4681 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4741 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4801 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4861 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4921 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4981 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5041 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5101 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5161 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     5221 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     5281 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     5341 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     5401 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     5461 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     5521 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     5581 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     5641 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     5701 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     5761 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     5821 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     5881 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     5941 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
     6001 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
     6061 ccaagctgga gacc
//
