LOCUS       40924_44878        4584 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC-gfp-neo, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4584)
  AUTHORS   Liu M, Zhang J.
  TITLE     Construction of GFP-Neo fusion protein by selection from a peptide 
            library
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4584)
  AUTHORS   Liu M, Zhang J.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-APR-2004) Microbiology, Immunology and Molecular 
            Genetics, University of Kentucky, 800 Rose Street, Lexington, KY 
            40536-0096, USA
REFERENCE   3  (bases 1 to 4584)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4584)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-APR-2004) Microbiology, Immunology and Molecular Genetics, 
            University of Kentucky, 800 Rose Street, Lexington, KY 40536-0096, 
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4584
                     /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"
     misc_feature    234..286
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             283..999
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1006..1071
                     /label=peptide linker between gfp and neo
                     /note="peptide linker between gfp and neo"
     CDS             1072..1863
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        2682..2786
                     /label=AmpR promoter
     CDS             2787..3644
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3818..4406
                     /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 gccaagcttg
      241 catgcctgca ggtcgactct agaggatcca ccggtcgcca ccatggtgag caagggcgag
      301 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      361 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      421 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      481 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      541 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      601 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      661 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      721 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
      781 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
      841 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
      901 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      961 gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgagtt
     1021 gcttattgtt cttctgatat gattgtgccc ttggatgagg ttggagctca tatgattgaa
     1081 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     1141 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     1201 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     1261 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     1321 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     1381 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     1441 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     1501 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     1561 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     1621 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     1681 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     1741 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     1801 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     1861 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     1921 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     1981 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccaccccg
     2041 ggctcgatcc cctcgcgagt tggttcagct gctgcctgag gctggacgac ctcgcggagt
     2101 tctaccggca gtgcaaatcc gtcggcatcc aggaaaccag cagcggctat ccgcgcatcc
     2161 atgcccccga actgcaggag tggggaggca cgatggccgc tttggtcgag gcggatctag
     2221 cttggggctg caggtcgact cactataggg agaccggaat tccaactgag cgccggtcgc
     2281 taccattacc aacttgtctg gtgtcaaaaa taataggcct actagtcggc cgtacgggcc
     2341 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2401 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2461 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     2521 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     2581 caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     2641 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2701 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2761 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2821 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2881 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2941 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3001 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3061 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3121 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3181 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3241 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3301 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3361 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3421 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3481 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3541 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3601 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3661 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3721 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3781 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3841 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3901 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3961 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4021 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4081 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4141 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4201 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4261 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4321 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4381 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4441 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4501 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4561 gcgagtcagt gagcgaggaa gcgg
//
