LOCUS       sequence-456516-        6585 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6585)
  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 6585)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pFLAG-HHARI
FEATURES             Location/Qualifiers
     source          1..6585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             618..641
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2609..3231
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2808..2827)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3255..3275)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3260..3589
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3440..3575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3627..3646)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3628..3646)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3660..3676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3660..3676)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3673..3695)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3684..3700
                     /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(3708..3714,3715..3732,3733..3738))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3753..3774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3891..3908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4062..4650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4142..4161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4821..5612,5613..5681))
                     /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     5444..5463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5682..5786)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5813..6268
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5900..5919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6110..6131
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6394..6416
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6408..6425
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6409..6425
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgatc taccatggac tacaaagacg atgacgacaa gcttatggac tcggacgagg
      661 gctacaacta cgagttcgac gaggacgagg agtgcagtga ggaggacagc ggcgccgagg
      721 aggaggagga cgaagacgac gacgagccgg acgatgatac cctggatctg ggcgaggtgg
      781 agctggtgga gcccgggctg ggcgtcggcg gggagcggga cggactgctg tgcggggaga
      841 cgggcggtgg cggcggcagc gctctggggc ccggcggtgg cggcggcggc ggcggcggcg
      901 gtggtggtgg cgggccgggg catgagcagg aggaggatta ccgctacgag gtgctcacgg
      961 ccgagcagat tctacaacac atggtggaat gtatccggga ggtcaacgag gtcatccaga
     1021 atccagcaac tatcacaaga atactcctta gccacttcaa ttgggataaa gagaagctaa
     1081 tggaaaggta ctttgatgga aacctggaga agctctttgc tgagtgtcat gtaattaatc
     1141 caagtaaaaa gtctcgaaca cgccagatga atacaaggtc atcagcacag gatatgcctt
     1201 gtcagatctg ctacttgaac taccctaact cgtatttcac tggccttgaa tgtggacata
     1261 agttttgtat gcagtgctgg agtgaatatt taactaccaa aataatggaa gaaggcatgg
     1321 gtcagactat ttcgtgtcct gctcatggtt gtgatatctt agtggatgac aacacagtta
     1381 tgcgcctgat cacagattca aaagttaaat taaagtatca gcatttaata acaaatagct
     1441 ttgtagagtg caatcgactg ttaaagtggt gtcctgcccc agattgccac catgttgtta
     1501 aagtccaata tcctgatgct aaacctgttc gctgcaaatg tgggcgccaa ttttgcttta
     1561 actgtggaga aaattggcat gatcctgtta aatgtaagtg gttaaagaaa tggattaaaa
     1621 agtgtgatga tgacagtgaa acctccaatt ggattgcagc caacacaaag gaatgtccca
     1681 aatgccatgt cacaattgag aaggatggtg gttgtaatca catggtctgt cgtaaccaga
     1741 attgtaaagc agagttttgc tgggtgtgtc ttggcccatg ggaaccacat ggatctgcct
     1801 ggtacaactg taaccgctat aatgaggatg atgcaaaggc agcaagagat gcacaggagc
     1861 gatctagggc agccctgcag aggtacctgt tctactgtaa tcgctatatg aaccacatgc
     1921 agagcctgcg ctttgagcac aaactatatg ctcaggtgaa acagaaaatg gaggagatgc
     1981 agcagcacaa catgtcctgg attgaggtgc agttcctgaa gaaggcagtt gatgtcctct
     2041 gccagtgtcg tgccacactc atgtacactt atgtcttcgc tttctacctc aaaaagaata
     2101 accagtccat tatctttgag aataaccaag cagatctaga gaatgccaca gaggtgctct
     2161 cgggctacct tgaacgagat atttcccaag attctctgca ggatataaag cagaaagtac
     2221 aagacaagta cagatactgt gagagtcgac gaagggtttt gttacagcat gtgcatgaag
     2281 gctatgaaaa agatctgtgg gagtacattg aggactgaga atggccctgc ataaaatgaa
     2341 ctctgaaaac tttaccatct agagtgctca tgcaattaaa acaaaacaaa cacaaacaag
     2401 gaggcactaa gcctattctg acaccactgg tctgtagtac cagaattgtt ttgttaatgg
     2461 aaagtttaag taaattatat tgtaataaaa aggtagataa acaaaaaaaa aaaaaaaaaa
     2521 aaaaaaaaaa aaaaaaaaaa aaaaaaaagg gcggccgcga attcatcgat agatctgata
     2581 tcggtaccag tcgactctag aggatcccgg gtggcatccc tgtgacccct ccccagtgcc
     2641 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     2701 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     2761 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     2821 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     2881 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     2941 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     3001 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     3061 aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc aggaggacgt
     3121 ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga gttgcttgct
     3181 tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat tgggtacgcg
     3241 gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3301 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3361 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3421 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3481 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3541 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcctcga
     3601 ggaactgaaa aaccagaaag ttaattccct atagtgagtc gtattaaatt cgtaatcatg
     3661 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3721 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3781 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3841 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3901 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3961 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4021 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4081 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4141 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4201 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4261 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4321 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4381 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4441 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4501 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4561 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4621 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4681 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4741 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4801 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4861 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4921 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4981 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5041 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5101 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5161 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5221 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5281 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5341 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5401 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5461 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5521 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5581 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5641 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5701 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5761 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
     5821 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5881 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5941 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6001 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6061 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6121 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     6181 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6241 tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc aactgttggg
     6301 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     6361 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     6421 ccagtgccaa gctgatctat acattgaatc aatattggca attagccata ttagtcattg
     6481 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     6541 atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgtt
//
