LOCUS       V043673                 6164 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043673
VERSION     V043673
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..1179
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     promoter        1196..1214
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     protein_bind    1293..1317
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             1348..2064
                     /label="Tnfsf14(NM_019418)"
                     /note="Tnfsf14(NM_019418)"
                     /gene="Tnfsf14"
     polyA_signal    2092..2316
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    2386..2419
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
     primer_bind     complement(2877..2893)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        3367..3471
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             3472..4332
                     /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"
     rep_origin      4503..5091
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    5379..5400
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5415..5445
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5453..5469
                     /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"
     primer_bind     5477..5493
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5991..6024
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctccg
     1321 cggccgcccc cttcaccatg ggccaccatg gagagtgtgg tacagccttc agtgtttgtg
     1381 gtggatggac agacggacat cccattcagg cggctggaac agaaccaccg gagacggcgc
     1441 tgtggcactg tccaggtcag cctggccctg gtgctgctgc taggtgctgg gctggccact
     1501 cagggctggt ttctcctgag actgcatcaa cgtcttggag acatagtagc tcatctgcca
     1561 gatggaggca aaggctcctg ggagaagctg atacaagatc aacgatctca ccaggccaac
     1621 ccagcagcac atcttacagg agccaacgcc agcttgatag gtattggtgg acctctgtta
     1681 tgggagacac gacttggcct ggccttcttg aggggcttga cgtatcatga tggggccctg
     1741 gtgaccatgg agcccggtta ctactatgtg tactccaaag tgcagctgag cggcgtgggc
     1801 tgcccccagg ggctggccaa tggcctcccc atcacccatg gactatacaa gcgcacatcc
     1861 cgctacccga aggagttaga actgctggtc agtcggcggt caccctgtgg ccgggccaac
     1921 agctcccgag tctggtggga cagcagcttc ctgggcggcg tggtacatct ggaggctggg
     1981 gaagaggtgg tggtccgcgt gcctggaaac cgcctggtca gaccacgtga cggcaccagg
     2041 tcctatttcg gagctttcat ggtctgattt aaacccgctg atcagcctcg actgtgcctt
     2101 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2161 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2221 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2281 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     2341 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaaataac ttcgtataat
     2401 gtatgctata cgaagttata tcgtatgcat gctttgcata cttctgcctg ctggggagcc
     2461 tggggacttt ccacacccta actgacacac attccacaga attccttgaa atacatccac
     2521 aggtacacct ccaattgact caaatgatgt caattagtct atcagaagct tctaaagcca
     2581 tgacatcatt ttctggaatt ttccaagctg tttaaaggca cagtcaactt agtgtatgta
     2641 aacttctgac ccactggaat tgtgatacag tgaattataa gtgaaataat ctgtctgtaa
     2701 acaattgttg gaaaaatgac ttgtgtcatg cacaaagtag atgtcctaac tgacttgcca
     2761 aaactattgt ttgttaacaa gaaatttgtg gagtagttga aaaacgagtt ttaatgactc
     2821 caacttaagt gtatgtaaac ttccgacttc aactgtaggg taccgagctc gaattcactg
     2881 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2941 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     3001 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     3061 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     3121 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3181 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3241 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3301 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3361 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3421 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3481 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3541 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3601 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3661 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3721 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3781 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3841 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3901 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3961 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4021 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4081 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4141 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4201 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4261 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4321 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4381 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4441 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4501 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4561 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4621 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4681 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4741 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4801 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4861 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4921 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4981 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5041 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5101 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5161 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5221 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     5281 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5341 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     5401 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     5461 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcctg
     5521 caggtcgact ctagacctat acagttgaag tcggaagttt acatacactt aagttggagt
     5581 cattaaaact cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc
     5641 aagtcagtta ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac
     5701 agacagatta tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca
     5761 tacactaagt tgactgtgcc tttaaacagc ttggaaaatt ccagaaaatg atgtcatggc
     5821 tttagaagct tctgatagac taattgacat catttgagtc aattggaggt gtacctgtgg
     5881 atgtatttca aggggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     5941 ctagaatgca gcggccgcta gctcgagatc ttaattaaac tagtatcgat ataacttcgt
     6001 ataatgtatg ctatacgaag ttatatgtac gggccagata tacgcgttga cattgattat
     6061 tgactaggct tttgcaaaaa gctttgcaaa gatggataaa gttttaaaca gagaggaatc
     6121 tttgcagcta atggaccttc taggtcttga aaggagtggg aatt
//
