LOCUS       sequence-485073-        6290 bp    DNA     circular SYN 29-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  AUTHORS   Nie M, Htun H
  TITLE     Different modes and potencies of translational repression by 
            sequence-specific RNA-protein interaction at the 5'-UTR.
  JOURNAL   Nucleic Acids Res. 2006  . 34(19):5528-40.
  PUBMED    17023487
REFERENCE   2  (bases 1 to 6290)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 29, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2006  . 34(19):5528-40."
COMMENT     Sequence Label: pIRE YIC
FEATURES             Location/Qualifiers
     source          1..6290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..80
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(105..209)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      236..691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(323..342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     533..554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    698..819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(709..728)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     763..782
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(959..976)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(980..1006)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(join(1025..1735,1736..1738,1739..1741))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    join(1742..1750,1751..1753,1754..2328)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1915..1934)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2144..2161
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(2381..2398)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2402..2425)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(join(2432..3142,3143..3145))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(3152..3181)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3188..3217)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3224..3253)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3329..3532)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3358..3378)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3533..3836
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4011..4599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4091..4110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4928..4975
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5000..5019
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5207..6001)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5207..6001)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5318..5337)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5928..5947
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6036..6290,1..103))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6050..6185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        join(6202..6290,1..103)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
ORIGIN
        1 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
       61 ccctaactga cacacattcc acagctggtt ctttccgcct caggactctt cctttttcaa
      121 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
      181 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
      241 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
      301 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
      361 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
      421 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
      481 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
      541 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
      601 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
      661 aattttaaca aaatattaac gcttacaatt tacgccttaa gatacattga tgagtttgga
      721 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
      781 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
      841 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
      901 aaatgtggta tggctgatta tgatctagag tcgcggccgc ttagttagtc agtcaggcgt
      961 gatggtggtg atggtgtcca gcgtaatctg gcacgtcgta tggatacagc tggtcgactc
     1021 tagacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     1081 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag ggcggactgg gtgctcaggt
     1141 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     1201 cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttg gccttgatgc
     1261 cgttcttctg cttgtcggcg gtgatataga cgttgtggct gatgtagttg tactccagct
     1321 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca
     1381 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga
     1441 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct
     1501 gcttcatgtg gtcggggtag cggctgaagc actgcacgcc ccaggtcagg gtggtcacga
     1561 gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc
     1621 cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc
     1681 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca
     1741 tggttgtggc catattatca tcgtgttttt caaaggaaaa ccacgtcccc gtggttcggg
     1801 gggcctagac gtttttttaa cctcgactaa acacatgtaa agcatgtgta ccgaggcccc
     1861 agatcagatc ccatacaatg gggtaccttc tgggcatcct tcagcccctt gttgaatacg
     1921 cttgaggaga gccatttgac tctttccaca actatccaac tcacaacgtg gcactggggt
     1981 tgtgccgcct ttgcaggtgt atcttataca cgtggctttt ggccgcagag gcacctgtcg
     2041 ccaggtgggg ggttccgctg cctgcaaagg gtcgctacag acgttgtttg tcttcaagaa
     2101 gcttccagag gaactgcttc cttcacgaca ttcaacagac cttgcattcc tttggcgaga
     2161 ggggaaagac ccctaggaat gctcgtcaag aagacagggc caggtttccg ggccctcaca
     2221 ttgccaaaag acggcaatat ggtggaaaat aacatataga caaacgcaca ccggccttat
     2281 tccaagcggc ttcggccagt aacgttaggg gggggggagg gagaggggcg gatcccgggc
     2341 ccgcggtacc gtcgactgca gaattcttag tcagtcaggc gtgatggtgg tgatggtggg
     2401 ccttatcgtc gtcatccttg taatcctcga gcttgtacag ctcgtccatg ccgagagtga
     2461 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     2521 tgctcagggc ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     2581 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
     2641 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
     2701 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     2761 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     2821 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     2881 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact
     2941 gcaggccgta gccgaaggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     3001 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     3061 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     3121 tgaacagctc ctcgcccttg ctcacgcgcg caagatcctc ctcggatatt aacttctgct
     3181 caccgttgag gtcttcctca ctgatcaatt tctgttctcc attcaagtcc tcttcagaaa
     3241 tgagcttttg ctccatggtg agctcgggtt ccgtccaagc actgttgaag caggaaaccc
     3301 gctagcggat ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac
     3361 gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt
     3421 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     3481 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa
     3541 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     3601 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     3661 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     3721 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     3781 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     3841 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactaatgc
     3901 atggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3961 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4021 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4081 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4141 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4201 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4261 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4321 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4381 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4441 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4501 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4561 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4621 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4681 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4741 agtatatatg agtaacctga ggctatggca gggcctgccg ccccgacgtt ggctgcgagc
     4801 cctgggcctt cacccgaact tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat
     4861 tggccccaat ggggtctcgg tggggtatcg acagagtgcc agccctggga ccgaaccccg
     4921 cgtttatgaa caaacgaccc aacaccgtgc gttttattct gtctttttat tgccgtcata
     4981 gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc cctagggtgg
     5041 gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa
     5101 acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc
     5161 aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt
     5221 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
     5281 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
     5341 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
     5401 cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc
     5461 cgtcgggcat gctcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
     5521 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
     5581 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
     5641 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
     5701 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
     5761 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt
     5821 gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
     5881 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
     5941 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca
     6001 tgcgaaacga tcctcatcct gtctcttgat cgatctttgc aaaagcctag gcctccaaaa
     6061 aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa
     6121 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     6181 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     6241 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
//
