LOCUS       V036960                 7089 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036960
VERSION     V036960
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1769
                     /label="SLC25A37(NM_016612)"
                     /note="SLC25A37(NM_016612)"
                     /gene="SLC25A37"
     CDS             1776..1841
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1865..2453
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2519..2567
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2769..3197
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3211..3540
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3607..4401
                     /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    4577..4698
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4747..4763)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4771..4787
                     /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(4795..4825)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4840..4861
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5149..5737)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5908..6768)
                     /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(6769..6873)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga gctgcgcagc gggagcgtgg
      781 gcagccaggc ggtggcgcgg aggatggatg gggacagccg agatggcggc ggcggcaagg
      841 acgccaccgg gtcggaggac tacgagaacc tgccgactag cgcctccgtg tccacccaca
      901 tgacagcagg agcgatggcc gggatcctgg agcactcggt catgtacccg gtggactcgg
      961 tgaagacacg aatgcagagt ttgagtccag atcccaaagc ccagtacaca agtatctacg
     1021 gagccctcaa gaaaatcatg cggaccgaag gcttctggag gcccttgcga ggcgtcaacg
     1081 tcatgatcat gggtgcaggg ccggcccatg ccatgtattt tgcctgctat gaaaacatga
     1141 aaaggacttt aaatgacgtt ttccaccacc aaggaaacag ccacctagcc aacgggatag
     1201 ctgggagtat ggccaccctg ctccacgatg cggtaatgaa tccagcagaa gtggtgaagc
     1261 agcgcttgca gatgtacaac tcgcagcacc ggtcagcaat cagctgcatc cggacggtgt
     1321 ggaggaccga ggggttgggg gccttctacc ggagctacac cacgcagctg accatgaaca
     1381 tccccttcca gtccatccac ttcatcacct atgagttcct gcaggagcag gtcaaccccc
     1441 accggaccta caacccgcag tcccacatca tctcaggcgg gctggccggg gccctcgccg
     1501 cggccgccac gacccccctg gacgtctgta agacccttct gaacactcag gagaacgtgg
     1561 ccctctcgct ggccaacatc agcggccggc tgtcgggtat ggccaatgcc ttccggacgg
     1621 tgtaccagct caacggcctg gccggctact tcaaaggcat ccaggcgcgt gtcatctacc
     1681 agatgccctc caccgccatt tcttggtctg tctatgagtt cttcaagtac tttctcacca
     1741 agcgccagct ggaaaatcga gctccatacg gatccgacta caaagaccat gacggtgatt
     1801 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     1861 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1921 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1981 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2041 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2101 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2161 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2221 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     2281 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2341 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2401 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     2461 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     2521 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     2581 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     2641 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     2701 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     2761 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2821 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2881 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     2941 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3001 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3061 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3121 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     3181 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3241 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3301 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3361 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3421 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3481 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3541 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3601 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3661 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3721 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3781 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3841 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3901 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3961 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4021 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4081 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4141 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4201 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4261 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4321 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4381 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     4441 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4501 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4561 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4621 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4681 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4741 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4801 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4861 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4921 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4981 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5041 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5101 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5161 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5221 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5281 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5341 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5401 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5461 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5521 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5581 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5641 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5701 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5761 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5821 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5881 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5941 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6001 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6061 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6121 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6181 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6241 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6301 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6361 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6421 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6481 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6541 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6601 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6661 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6721 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6781 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6841 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6901 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     6961 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7021 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     7081 ctgcttagg
//
