LOCUS       40924_33277        4504 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Bacterial expression vector pNIL2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4504)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4504)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 4504)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4504)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      27..75
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     regulatory      134..215
                     /label=fd terminator
                     /note="fd terminator"
                     /regulatory_class="terminator"
     terminator      137..185
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      313..768
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(927..1515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1689..2546)
                     /label=AmpR
                     /note="beta-lactamase"
     misc_feature    complement(2861..3520)
                     /label=chloramphenicol sensitive
                     /note="chloramphenicol sensitive"
     CDS             complement(2861..3520)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTV*LDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3521..3623)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     promoter        3788..3816
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    3824..3840
                     /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)."
     misc_feature    3865..3881
                     /label=5' UTR of ompA, incl. RBS
                     /note="5' UTR of ompA, incl. RBS"
     sig_peptide     3882..3944
                     /label=OmpA signal peptide
                     /note="signal peptide from the E. coli outer membrane
                     protein OmpA"
     CDS             3945..4394
                     /codon_start=1
                     /note="unnamed protein product; mIL2 mature sequence"
                     /protein_id="SJL88449.1"
                     /translation="APTSSSTSSSTAEAQQQQQQQQQQQQHLEQLLMDLQELLSRMENY
                     RNLKLPRMLTFKFYLPKQATELKDLQCLEDELGPLRHVLDLTQSKSFQLEDAENFISNI
                     RVTVVKLKGSDNTFECQFDDESATVVDFLRRWIAFCQSIISTSPQ"
     misc_feature    4395..4497
                     /label=3' UTR of mIL2
                     /note="3' UTR of mIL2"
ORIGIN
        1 aattcacctc gaaagcaagc tgataaaccg atacaattaa aggctccttt tggagccttt
       61 ttttttggag attttcaacg tgaaaaaatt attattcgca attccaagct aattcacctc
      121 gaaagcaagc tgataaaccg atacaattaa aggctccttt tggagccttt ttttttggag
      181 attttcaacg tgaaaaaatt attattcgca attccaagct ctgcctcgcg cgtttcggtg
      241 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
      301 cggatgcaga tcacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
      361 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
      421 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
      481 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
      541 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
      601 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
      661 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
      721 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttga tctgcgctcg
      781 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      841 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
      901 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      961 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1021 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1081 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1141 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1201 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1261 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1321 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     1381 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1441 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1501 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1561 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1621 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1681 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1741 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1801 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     1861 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     1921 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     1981 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2041 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2101 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2161 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2221 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2281 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     2341 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2401 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     2461 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     2521 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcagacag
     2581 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     2641 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgactcccc gcgcgcgatg
     2701 ggtcgaattt gctttcgaaa aaaaagcccg ctcattaggc gggctaaaaa aaagcccgct
     2761 cattaggcgg gctcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     2821 aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact
     2881 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacagacgg
     2941 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     3001 ccatagtgaa aacgggggcg aagaagttgt ccatattcgc cacgtttaaa tcaaaactgg
     3061 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     3121 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     3181 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     3241 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     3301 gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     3361 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctaa acggtctggt
     3421 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     3481 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     3541 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     3601 tggaacctct tacgtgccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc
     3661 cggtatcaac agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta
     3721 tttattcgaa gacgaaaggg catcgcgcgc ggggaattcg agctcgagct tactccccat
     3781 ccccctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa
     3841 tttcacacag gaaacaggat cctagataac gagggcaaaa aatgaaaaag acagctatcg
     3901 cgattgcagt ggcactggct ggtttcgcta ccgtagcgca ggccgcaccc acttcaagct
     3961 ccacttcaag ctctacagcg gaagcacagc agcagcagca gcagcagcag cagcagcagc
     4021 agcacctgga gcagctgttg atggacctac aggagctcct gagcaggatg gagaattaca
     4081 ggaacctgaa actccccagg atgctcacct tcaaatttta cttgcccaag caggccacag
     4141 aattgaaaga tcttcagtgc ctagaagatg aacttggacc tctgcggcat gttctggatt
     4201 tgactcaaag caaaagcttt caattggaag atgctgagaa tttcatcagc aatatcagag
     4261 taactgttgt aaaactaaag ggctctgaca acacatttga gtgccaattc gatgatgagt
     4321 cagcaactgt ggtggacttt ctgaggagat ggatagcctt ctgtcaaagc atcatctcaa
     4381 caagccctca ataactatgt acctcctgct tacaacacat aaggctctct atttatttaa
     4441 atatttaact ttaatttatt tttggatgta ttgtttacta tcttttgtaa ctactagagg
     4501 tcga
//
