LOCUS       V040855                 7753 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040855
VERSION     V040855
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             928..3189
                     /label="RXFP2(NM_130806)"
                     /note="RXFP2(NM_130806)"
                     /gene="RXFP2"
     CDS             3196..3219
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             3226..3315
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     polyA_signal    3353..3577
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3623..4051
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4065..4394
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4461..5255
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5429..5550
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5599..5615)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5623..5639
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5647..5677)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5692..5713
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6001..6586)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6757..7617)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7618..7722)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagctgaatt cgccaccatg attgtttttc tggtttttaa acatctcttc
      961 agcctcagat tgattacaat gttctttcta cttcatttca tcgttctgat caatgtcaaa
     1021 gattttgcac tgactcaagg tagcatgatc actccttcat gccaaaaagg atattttccc
     1081 tgtgggaatc ttaccaagtg cttaccccga gcttttcact gtgatggcaa ggatgactgt
     1141 gggaacgggg cggacgaaga gaactgtggt gacactagtg gatgggcgac catatttggc
     1201 acagtgcatg gaaatgctaa cagcgtggcc ttaacacagg agtgctttct aaaacagtat
     1261 ccacaatgct gtgactgcaa agaaactgaa ttggaatgtg taaatggtga cttaaagtct
     1321 gtgccgatga tttctaacaa tgtgacatta ctgtctctta agaaaaacaa aatccacagt
     1381 cttccagata aagttttcat caaatacaca aaacttaaaa agatatttct tcagcataat
     1441 tgcattagac acatatccag gaaagcattt tttggattat gtaatctgca aatattatat
     1501 ctcaaccaca actgcatcac aaccctcaga cctggaatat tcaaagactt acatcagcta
     1561 acttggctaa ttctagatga caatccaata accagaattt cacagcgctt gtttacggga
     1621 ttaaattcct tgtttttcct gtctatggtt aataactact tagaagctct tcccaagcag
     1681 atgtgtgccc aaatgcctca actcaactgg gtggatttgg aaggcaatag aataaagtat
     1741 ctcacaaatt ctacgtttct gtcgtgcgat tcgctcacag tgctgtttct gcctagaaat
     1801 caaattggtt ttgttccaga gaagacattt tcttcattaa aaaatttagg agaactggat
     1861 ctgtctagca atacgataac ggaactatca cctcaccttt ttaaagactt gaagcttcta
     1921 caaaagctga acctgtcatc caatcctctt atgtatcttc acaagaacca gtttgaaagt
     1981 cttaaacaac ttcagtctct agacctggaa aggatagaga ttccaaatat aaacacacga
     2041 atgtttcaac ccatgaagaa tctttctcac atttatttca aaaactttcg atactgctcc
     2101 tatgctcccc atgtccgaat atgtatgccc ttgacggacg gcatttcttc atttgaggac
     2161 ctcttggcta acaatatcct cagaatattt gtctgggtta tagctttcat tacctgcttt
     2221 ggaaatcttt ttgtcattgg catgagatct ttcattaaag ctgaaaatac aactcacgct
     2281 atgtccatca aaatcctttg ttgtgctgat tgcctgatgg gtgtttactt gttctttgtt
     2341 ggcattttcg atataaaata ccgagggcag tatcagaagt atgccttgct gtggatggag
     2401 agcgtgcagt gccgcctcat ggggttcctg gccatgctgt ccaccgaagt ctctgttctg
     2461 ctactgacct acttgacttt ggagaagttc ctggtcattg tcttcccctt cagtaacatt
     2521 cgacctggaa aacggcagac ctcagtcatc ctcatttgca tctggatggc gggattttta
     2581 atagctgtaa ttccattttg gaataaggat tattttggaa acttttatgg gaaaaatgga
     2641 gtatgtttcc cactttatta tgaccaaaca gaagatattg gaagcaaagg gtattctctt
     2701 ggaattttcc taggtgtgaa cttgctggct tttctcatca ttgtgttttc ctatattact
     2761 atgttctgtt ccattcaaaa aaccgccttg cagaccacag aagtaaggaa ttgttttgga
     2821 agagaggtgg ctgttgcaaa tcgtttcttt tttatagtgt tctctgatgc catctgctgg
     2881 attcctgtat ttgtagttaa aatcctttcc ctcttccggg tggaaatacc agacacaatg
     2941 acttcctgga tagtgatttt tttccttcca gttaacagtg ctttgaatcc aatcctctat
     3001 actctcacaa ccaacttttt taaggacaag ttgaaacagc tgctgcacaa acatcagagg
     3061 aaatcaattt tcaaaattaa aaaaaaaagt ttatctacat ccattgtgtg gatagaggac
     3121 tcctcttccc tgaaacttgg ggttttgaac aaaataacac ttggagacag tataatgaaa
     3181 ccagtttccc tcgaggatta caaggatgac gacgataagg gatccagcgc ctggtcccac
     3241 cctcagtttg agaagggcgg aggctctggc ggcggaagcg gaggatctgc ttggagccac
     3301 ccccagttcg aaaagtgatg agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3361 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3421 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3481 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3541 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3601 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3661 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3721 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3781 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3841 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3901 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3961 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4021 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4081 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4141 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4201 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4261 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4321 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4381 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4441 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4501 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4561 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4621 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4681 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4741 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4801 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4861 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4921 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4981 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5041 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5101 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5161 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5221 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5281 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5341 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5401 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5461 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5521 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5581 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5641 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5701 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5761 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5821 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5881 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5941 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6001 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6061 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6541 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6601 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6661 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6721 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6781 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6841 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6901 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6961 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7021 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7081 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7141 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7201 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7261 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7321 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7381 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7441 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7501 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7561 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7621 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7681 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7741 gccacctgac gtc
//
