LOCUS       40924_16785        6007 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pECIA-2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6007)
  AUTHORS   Ozkan E, Carrillo RA, Eastman CL, Weiszmann R, Waghray D, Johnson 
            KG, Zinn K, Celniker SE, Garcia KC.
  TITLE     An Extracellular Interactome of Immunoglobulin and LRR Proteins 
            Reveals Receptor-Ligand Networks
  JOURNAL   Cell 154 (1), 228-239 (2013)
  PUBMED    23827685
REFERENCE   2  (bases 1 to 6007)
  AUTHORS   Ozkan E, Eastman CL, Garcia KC.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JUL-2013) Molecular and Cellular Physiology, Stanford 
            University School of Medicine, 279 Campus Drive, Beckman Center, 
            Stanford, CA 94306, USA
REFERENCE   3  (bases 1 to 6007)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6007)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell"; 
            date: "2013"; volume: "154"; issue: "1"; pages: "228-239"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-JUL-2013) Molecular and Cellular Physiology, Stanford University
            School of Medicine, 279 Campus Drive, Beckman Center, Stanford, CA 
            94306, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        401..827
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     845..898
                     /label=BiP signal sequence
                     /note="Drosophila BiP signal sequence"
     protein_bind    908..1032
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1057..1087
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1141..1797
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2142..2444
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2488..2612)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2638..2661
                     /codon_start=1
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
                     /translation="LEVLFQGP"
     misc_feature    2668..3348
                     /label=IgG1 heavy chain Fc fragment
                     /note="IgG1 heavy chain Fc fragment"
     CDS             3367..3408
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             3418..3435
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    complement(3508..3642)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3786..3802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3810..3826)
                     /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)."
     promoter        complement(3834..3864)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3879..3900)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4188..4776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4950..5807)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5808..5912)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt aattcgttgc aggacaggat
      421 gtggtgcccg atgtgactag ctctttgctg caggccgtcc tatcctctgg ttccgataag
      481 agacccagaa ctccggcccc ccaccgccca ccgccacccc catacatatg tggtacgcaa
      541 gtaagagtgc ctgcgcatgc cccatgtgcc ccaccaagag ctttgcatcc catacaagtc
      601 cccaaagtgg agaaccgaac caattcttcg cgggcagaac aaaagcttct gcacacgtct
      661 ccactcgaat ttggagccgg ccggcgtgtg caaaagaggt gaatcgaacg aaagacccgt
      721 gtgtaaagcc gcgtttccaa aatgtataaa accgagagca tctggccaat gtgcatcagt
      781 tgtggtcagc agcaaaatca agtgaatcat ctcagtgcaa ctaaaggggg gatccgatct
      841 caatatgaag ttatgcatat tactggccgt cgtggccttt gttggcctct cgctcgggag
      901 atcatcaaca agtttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa
      961 tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat
     1021 ccagtcacta tggcggccgc attaggcacc ccaggcttta cactttatgc ttccggctcg
     1081 tataatgtgt ggattttgag ttaggatccg tcgagatttt caggagctaa ggaagctaaa
     1141 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     1201 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     1261 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     1321 cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat gaaagacggt
     1381 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     1441 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     1501 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     1561 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     1621 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     1681 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
     1741 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
     1801 agatctggat ccggcttact aaaagccaga taacagtatg cgtatttgcg cgctgatttt
     1861 tgcggtataa gaatatatac tgatatgtat acccgaagta tgtcaaaaag aggtatgcta
     1921 tgaagcagcg tattacagtg acagttgaca gcgacagcta tcagttgctc aaggcatata
     1981 tgatgtcaat atctccggtc tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg
     2041 tgccgaacgc tggaaagcgg aaaatcagga agggatggct gaggtcgccc ggtttattga
     2101 aatgaacggc tcttttgctg acgagaacag gggctggtga aatgcagttt aaggtttaca
     2161 cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca
     2221 cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct
     2281 cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg
     2341 atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg
     2401 aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc
     2461 ttatacacag ccagtctgca ggtcgaccat agtgactgga tatgttgtgt tttacagtat
     2521 tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac
     2581 gtttctcgtt cagctttctt gtacaaagtg gttgatatcc agcacagtgg cggccgcttg
     2641 gaagttctgt ttcaagggcc tgggagcgac aaaactcaca catgcccacc gtgcccagca
     2701 cctgaactcc tggggggacc gtcagtcttc ctcttccccc caaaacccaa ggacaccctc
     2761 atgatctccc ggacccctga ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct
     2821 gaggtcaagt tcaactggta cgtggacggc gtggaggtgc ataatgccaa gacaaagccg
     2881 cgggaggagc agtacaacag cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag
     2941 gactggctga atggcaagga gtacaagtgc aaggtctcca acaaagccct cccagccccc
     3001 atcgagaaaa ccatctccaa agccaaaggg cagccccgag aaccacaggt gtacaccctg
     3061 cccccatccc gggaggagat gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc
     3121 ttctatccca gcgacatcgc cgtggagtgg gagagcaatg ggcagccgga gaacaactac
     3181 aagaccacgc ctcccgtgct ggactccgac ggctccttct tcctctacag caagctcacc
     3241 gtggacaaga gcaggtggca gcaggggaac gtcttctcat gctccgtgat gcatgagggt
     3301 ctgcacaacc actacacgca gaagagcctc tccctgtctc cgggtaaatc tagagggccc
     3361 ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg taccggtcat
     3421 catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc
     3481 tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat tgatgagttt
     3541 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3601 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3661 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     3721 tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa
     3781 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3841 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3901 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3961 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4021 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4081 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4141 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4201 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4261 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4321 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4381 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4441 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4501 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4561 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4621 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4681 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4741 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4801 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4861 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4921 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4981 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5041 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5101 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5161 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5221 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5281 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5341 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5401 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5461 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5521 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5581 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5641 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5701 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5761 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5821 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5881 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5941 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6001 tttcgtc
//
