LOCUS       40924_17094        8103 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEFBOS-IRESGFPNeo, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8103)
  AUTHORS   Lim SM, Stanley EG, Elefanty AG.
  TITLE     pEFBOS-IRESGFPNeo, a derivative of pEFBOS containing an IRESGFPNeo 
            cassette
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8103)
  AUTHORS   Lim SM, Stanley EG, Elefanty AG.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-FEB-2007) Monash Immunology and Stem Cell 
            Laboratories, Monash University, West Ring Road, Clayton, Vic 3800, 
            Australia
REFERENCE   3  (bases 1 to 8103)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8103)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-FEB-2007) Monash Immunology and Stem Cell Laboratories, Monash 
            University, West Ring Road, Clayton, Vic 3800, Australia"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(333..543)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        562..1743
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     misc_feature    1826..2412
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2425..3135
                     /codon_start=1
                     /label=mgfp5
                     /note="GFP with folding enhancement mutations"
                     /translation="SKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
                     FKTRHNIEDGGVQLADHYQQNTPIGDDPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             3145..3933
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELCGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     3'UTR           4523..5223
                     /label=bovine growth hormone 3' UTR
                     /note="bovine growth hormone 3' UTR"
     primer_bind     complement(5232..5248)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5461..5916
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6198..6302
                     /label=AmpR promoter
     CDS             6303..7160
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7334..7922
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttc
      241 caggcactcc tttcaagacc tagaaggtcc attagctgca aagattcctc tctgtttaaa
      301 actttatcca tctttgcaaa gctttttgca aaagcctagg cctccaaaaa agcctcctca
      361 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
      421 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
      481 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
      541 tgctggggag cctggcgtga ggctccggtg cccgtcagtg ggcagagcgc acatcgccca
      601 cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc
      661 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
      721 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
      781 ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg
      841 gcccttgcgt gccttgaatt acttccacgc ccctggctgc agtacgtgat tcttgatccc
      901 gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg
      961 cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
     1021 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
     1081 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac
     1141 actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca
     1201 tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa
     1261 gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg
     1321 gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct
     1381 gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc
     1441 acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac
     1501 cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt
     1561 tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt
     1621 aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct
     1681 tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg
     1741 tgaggaattc tctaggtcga ccccggggat cctctagact agtgctagca tgctgcagtc
     1801 gaaggtaccg cgggcccggg atccgcccct ctccctcccc cccccctaac gttactggcc
     1861 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     1921 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     1981 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     2041 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     2101 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     2161 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     2221 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     2281 tgggatctga tctggggcct cggtacacat gctttacatg tgtttagtcg aggttaaaaa
     2341 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     2401 atggccacaa ccatggcgcg ccagagtaaa ggagaagaac ttttcactgg agttgtccca
     2461 attcttgttg aattagatgg tgatgttaat gggtacaaat tttctgtcag tggagagggt
     2521 gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta
     2581 cctgttccat ggccaacact tgtcactact ctcacttatg gtgttcaatg cttttcaaga
     2641 tatccagatc atatgaagcg gcacgacttc ttcaagagcg ccatgcctga gggatacgtg
     2701 caggagagga ccatcttctt caaggacgac gggaactaca agacacgtgc tgaagtcaag
     2761 tttgagggag acaccctcgt caacaggatc gagcttaagg gaatcgattt caaggaggac
     2821 ggaaacatcc tcggccacaa gttggaatac aactacaact cccacaacgt atacatcatg
     2881 gccgacaagc aaaagaacgg catcaaagcc aacttcaaga cccgccacaa catcgaagac
     2941 ggcggcgtgc aactcgctga tcattatcaa caaaatactc caattggcga tgaccctgtc
     3001 cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa
     3061 aagagagacc acatggtcct tcttgagttt gtaacggctg ctgggattac acatggcatg
     3121 gatgaactat acaaaacgcg tgggattgaa caagatggat tgcacgcagg ttctccggcc
     3181 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3241 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3301 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3361 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3421 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3481 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3541 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3601 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3661 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3721 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3781 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagctttgc
     3841 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3901 atcgccttct atcgccttct tgacgagttc ttctgaaaat tgatgatcta ttaaacaata
     3961 aagatgtcca ctaaaatgga agttttttac tgtcatactt tgttaagaag ggtgagaaca
     4021 gagtacctac attttgaatg gaaggattgg agctacgggg gtgggggtgg gggtgggatt
     4081 agataaatgc ctgctcttta ctgaaggctc tttactattg ctttatgata atgtttcata
     4141 gttggatatc ataatttaaa caagcaaaac caaattaagg gccagctcat tcctccactc
     4201 atgatctata gatctataga tctctcgggg gatcattgtt tttttcttga ttcccacttt
     4261 gtggttctaa gtactgtggt ttccaaatgt gtcagtttca tagcctgaag aacgagatca
     4321 gcagcctctg ttccacatac acttcattct cagtattgtt ttgccaagtt ctaattccat
     4381 cagaagctga ctctagagtg agggtcccca cctgggaccc ttgagagtat caggtctccc
     4441 acgtgggaga caagaaatcc ctgtttaata tttaaacagc agtgttcccc atctgggtcc
     4501 ttgcacccct cactctggcc tcagccgact gcacagcggc ccctgcatcc ccttggctgt
     4561 gaggcccctg gacaagcaga ggtggccaga gctgggaggc atggccctgg ggtcccacga
     4621 atttgctggg gaatctcgtt tttcttctta agacttttgg gacatggttt gactcccgaa
     4681 catcaccgac gtgtctcctg tttttctggg tggcctcggg acacctgccc tgcccccacg
     4741 agggtcagga ctgtgactct ttttagggcc aggcaggtgc ctggacattt gccttgctgg
     4801 atggggactg gggatgtggg agggagcaga caggaggaat catgtcaggc ctgtgtgtga
     4861 aaggaagctc cactgtcacc ctccacctct tcacccccca ctcaccagtg tcccctccac
     4921 tgtcacattg taactgaact tcaggataat aaagtgtttg cctccagtca cgtccttcct
     4981 ccttcttgag tccagctggt gcctggccag gggctgggga ggtggctgaa gggtgggaga
     5041 ggccagaggg aggtcgggga ggaggtctgg ggaggaggtc cagggaggag gaggaaagtt
     5101 ctcaagttcg tctgacattc attccgttag cacatattta tctgagcacc tactctgtgc
     5161 agacgctggg ctaagtgctg gggacacagc agggaacaag gcagacatgg aatctgcact
     5221 cganngaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     5281 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     5341 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     5401 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     5461 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     5521 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5581 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     5641 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     5701 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5761 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat
     5821 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     5881 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     5941 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     6001 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     6061 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     6121 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     6181 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6241 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6301 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6361 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6421 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6481 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6541 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6601 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6661 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6721 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6781 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6841 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6901 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6961 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     7021 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7081 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7141 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7201 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7261 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7321 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7381 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7441 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     7501 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7561 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7621 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7681 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7741 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7801 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7861 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7921 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7981 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     8041 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     8101 aag
//
