LOCUS       Exported                6339 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000531 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6339)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 2, 2021 from SnapGene 5.3.1
            https://www.snapgene.com
COMMENT     This file was created using tools provided by Twist Bioscience
FEATURES             Location/Qualifiers
     source          1..6339
                     /organism="BBF10K_000531 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..1845
                     /codon_start=1
                     /product="Nuclear Receptor Subfamily 4 Group A Member 2"
                     /label=NR4A2
                     /translation="MPCVQAQYGSSPQGASPASQSYSYHSSGEYSSDFLTPEFVKFSMD
                     LTNTEITATTSLPSFSTFMDNYSTGYDVKPPCLYQMPLSGQQSSIKVEDIQMHNYQQHS
                     HLPPQSEEMMPHSGSVYYKPSSPPTPTTPGFQVQHSPMWDDPGSLHNFHQNYVATTHMI
                     EQRKTPVSRLSLFSFKQSPPGTPVSSCQMRFDGPLHVPMNPEPAGSHHVVDGQTFAVPN
                     PIRKPASMGFPGLQIGHASQLLDTQVPSPPSRGSPSNEGLCAVCGDNAACQHYGVRTCE
                     GCKGFFKRTVQKNAKYVCLANKNCPVDKRRRNRCQYCRFQKCLAVGMVKEVVRTDSLKG
                     RRGRLPSKPKSPQEPSPPSPPVSLISALVRAHVDSNPAMTSLDYSRFQANPDYQMSGDD
                     TQHIQQFYDLLTGSMEIIRGWAEKIPGFADLPKADQDLLFESAFLELFVLRLAYRSNPV
                     EGKLIFCNGVVLHRLQCVRGFGEWIDSIVEFSSNLQNMNIDISAFSCIAALAMVTERHG
                     LKEPKRVEELQNKIVNCLKDHVTFNNGGLNRPNYLSKLLGKLPELRTLCTQGLQRIFYL
                     KLEDLVPPPAIIDKLFLDTLPF"
     primer_bind     complement(1846..1870)
                     /label=Gibson-rev
     terminator      1989..2036
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2041..2057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2216..2466
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(2469..3224)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      3315..3534
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    3954..3975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3990..4020
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4028..4044
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4052..4068
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4141..4986
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    5094..5245
                     /label=nnpp promoters
     CDS             5285..6100
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      complement(6210..6253)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     6271..6287
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6288..6319
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgccctgc
       61 gtgcaggccc agtacggcag cagcccccag ggcgccagcc ccgccagcca gagctacagc
      121 taccacagca gcggcgagta cagcagcgac ttcctgaccc ccgagttcgt gaagttcagc
      181 atggacctga ccaacaccga gatcaccgcc accaccagcc tgcccagctt cagcaccttc
      241 atggacaact acagcaccgg ctacgacgtg aagcccccct gcctgtacca gatgcccctg
      301 agcggccagc agagcagcat caaggtggag gacatccaga tgcacaacta ccagcagcac
      361 agccacctgc ccccccagag cgaggagatg atgccccaca gcggcagcgt gtactacaag
      421 cccagcagcc cccccacccc caccaccccc ggcttccagg tgcagcacag ccccatgtgg
      481 gacgaccccg gcagcctgca caacttccac cagaactacg tggccaccac ccacatgatc
      541 gagcagagaa agacccccgt gagcagactg agcctgttca gcttcaagca gagccccccc
      601 ggcacccccg tgagcagctg ccagatgaga ttcgacggcc ccctgcacgt gcccatgaac
      661 cccgagcccg ccggcagcca ccacgtggtg gacggccaga ccttcgccgt gcccaacccc
      721 atcagaaagc ccgccagcat gggcttcccc ggcctgcaga tcggccacgc cagccagctg
      781 ctggacaccc aggtgcccag cccccccagc agaggcagcc ccagcaacga gggcctgtgc
      841 gccgtgtgcg gcgacaacgc cgcctgccag cactacggcg tgagaacctg cgagggctgc
      901 aagggcttct tcaagagaac cgtgcagaag aacgccaagt acgtgtgcct ggccaacaag
      961 aactgccccg tggacaagag aagaagaaac agatgccagt actgcagatt ccagaagtgc
     1021 ctggccgtgg gcatggtgaa ggaggtggtg agaaccgaca gcctgaaggg cagaagaggc
     1081 agactgccca gcaagcccaa gagcccccag gagcccagcc cccccagccc ccccgtgagc
     1141 ctgatcagcg ccctggtgag agcccacgtg gacagcaacc ccgccatgac cagcctggac
     1201 tacagcagat tccaggccaa ccccgactac cagatgagcg gcgacgacac ccagcacatc
     1261 cagcagttct acgacctgct gaccggcagc atggagatca tcagaggctg ggccgagaag
     1321 atccccggct tcgccgacct gcccaaggcc gaccaggacc tgctgttcga gagcgccttc
     1381 ctggagctgt tcgtgctgag actggcctac agaagcaacc ccgtggaggg caagctgatc
     1441 ttctgcaacg gcgtggtgct gcacagactg cagtgcgtga gaggcttcgg cgagtggatc
     1501 gacagcatcg tggagttcag cagcaacctg cagaacatga acatcgacat cagcgccttc
     1561 agctgcatcg ccgccctggc catggtgacc gagagacacg gcctgaagga gcccaagaga
     1621 gtggaggagc tgcagaacaa gatcgtgaac tgcctgaagg accacgtgac cttcaacaac
     1681 ggcggcctga acagacccaa ctacctgagc aagctgctgg gcaagctgcc cgagctgaga
     1741 accctgtgca cccagggcct gcagagaatc ttctacctga agctggagga cctggtgccc
     1801 ccccccgcca tcatcgacaa gctgttcctg gacaccctgc ccttcggatc cgccaccaac
     1861 ttcagcctgc tagtattagg cgtcaaggtc catcttcctg accgaagtag tatagggata
     1921 acagggtaat tgccattgcg gaaatttttg gcgctgaggt gtcaatcgtc ggagccgctg
     1981 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgcatg
     2041 gtcatagctg tttcctgaga gcttggcagg tgtcgcgtat gcatcgatca cgtgtccacg
     2101 ttctttaata gtggactctt gttccaaact ggaacaacac tcggatcgat ccggcgcgca
     2161 ccgtgggaaa aactccaggt agaggtacac acgcggatag ccaattcaga gtaataaact
     2221 gtgataatca accctcatca atgatgacga actaaccccc gatatcaggt cacatgacga
     2281 agggaaagag aaggaaatca actgtgacaa actgccctca aatttggctt ccttaaaaat
     2341 tacagttcaa aaagtatgag aaaatccatg caggctgaag gaaacagcaa aactgtgaca
     2401 aattaccctc agtaggtcag aacaaatgtg acgaaccacc ctcaaatctg tgacagataa
     2461 ccctcagact atcctgtcgt catggaagtg atatcgcgga aggaaaatac gatatgagtc
     2521 gtctggcggc ctttcttttt ctcaatgtat gagaggcgca ttggagttct gctgttgatc
     2581 tcattaacac agacttgcag gaagcggcgg cggaagtcag gcatacgctg gtaactttga
     2641 ggcagctggt aacgctctat gatccagtcg attttcagag acacgatgcc tgagccatcc
     2701 ggcttacgat actgacacag ggattcgtat aaacgcatgg catacggatt ggtgatttct
     2761 tttgtttcac taagccgaaa ctgcgtaaac cggttctgta acccgataaa gaagggaatg
     2821 agatatgggt tgatatgtac actgtaaagc cctctggatg gactgtgcgc acgtttgata
     2881 aaccaaggaa aagattcata gcctttttca tcaccggcat cctcttcagg gcgataaaaa
     2941 accacttcct tccccgcgaa actcttcaat gcctgccgta tatccttact ggcttccgca
     3001 gaggtcaatc cgaatatttc agcatattta gcaacatgga tctcgcagat accgtcatgt
     3061 tcctgtaggg tgccatcaga ttttctgatc tggtcaacga acagatacag catacgtttt
     3121 tgatcccggg agagactata tgccgcctca gtgaggtcgt ttgactggac gattcgcggg
     3181 ctatttttac gtttcttgtg attgataacc gctgtttccg ccatgacaga tccatgtgaa
     3241 gtgtgacaag tttttagatt gtcacactaa ataaaaaaga gtcaataagc agggataact
     3301 ttgtgaaaaa acagcttctt ctgagggcaa tttgtcacag ggttaagggc aatttgtcac
     3361 agacaggact gtcatttgag ggtgatttgt cacactgaaa gggcaatttg tcacaacacc
     3421 ttctgtagaa ccagcatgga taaaggccta caaggcgctc taaaaaagaa gatctaaaaa
     3481 ctataaaaaa aataattata aaaatatccc cgtggataag tggataaccc caagggaagt
     3541 tttttcaggc atcgtgtgta agcagaatat ataagtgctg ttccctggtg cttcctcgct
     3601 cactcgaaat tcccggggat agctttatgc ttgtaaaccg ttttgtgaaa aaatttttaa
     3661 aataaaaaag gggacctcta gggtccccaa ttaattagta atataatcta ttaaaggtca
     3721 ttcaaaaggt catccaccgg atcaattccc ctgctcgcgc aggctgggtg ccaagctctc
     3781 gggtaacatc aaggcccgat ccttggagcc cttgccctcc cgcacgatga tcgtgccgtg
     3841 atcgaaatcc agatccttga cccgcagttg caaaccctca ctgatccgat tcattaatgc
     3901 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     3961 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     4021 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     4081 aagcgcggta cctgtcgcgg caacgcgcta acagacgtag taagaaccac cagcattgta
     4141 atgctggcta aagtcacttt cctgagctgt ataacgatga gcgattttac tttttctggc
     4201 tatgaattgg cctgctttgt aacacactcc ggtctatccc gtagcgccgg gcatatcctg
     4261 tcgcaatgtg caaatctcgc ggcaacaacc agtgaatact tcattcacaa gcctcaccgc
     4321 ctgatcgcgg cagaaactgg ttatagccaa tcaaccgtcg ttcgtgcatt ccgtgaagct
     4381 gtaaacaaag gtatcctgtc tgtagagatt gttatcggcg atcaccgtga acgtcgcgct
     4441 aacctgtacc ggtttacacc atcctttttg gccttcgcac aacaagccaa aaatgcgctg
     4501 atagaaagca aattaaagat ctcttcagcg gcaaccaagg ttaaagctgt tctcgctaag
     4561 acattggctt tatttaattt tttatccaca cccccatgtc aaaatgatac cccctccccc
     4621 tgtcaggatg acgtggcaat aaagaataag aagtcacaag ttaaaaaaac aaaaagatca
     4681 gtttccggcg gtgccggaac aaccagcctc aaaaaattga cttcatggat cgctaaggca
     4741 aaagcaaagg ctgacaatct gcggttatcc aaaaaacgca ctcaaaaaca tgagttcaag
     4801 cagaaagtag aggcggctgc gcggaaatat gcttacctga agaacaagcg ttcgcctgat
     4861 attggcggga tatcaaactt cgataaccta ccgcattgca tgacggtaaa cgaagctctt
     4921 aatgcggttt tagccaaaaa taaagataac gaacaatggg gtataccggc aggattcaga
     4981 gggtaatgaa ttgctctaat tataaccatg catactttca acacctctag tttgccatga
     5041 ggcaaactca taggtgtcct ggtaagagga cactgttgcc aaaactggac gccccattat
     5101 tgcaattaat aaacaactaa cggacaattc tacctaacaa taagtggagt tgcggccggc
     5161 cgcgatccgc tagcaaagcc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     5221 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     5281 tgttatgagc catattcaac gggaaacgtc ttgctcgcgc ccgcgattaa attccaacat
     5341 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     5401 aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     5461 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     5521 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     5581 tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     5641 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     5701 tccttttaac agcgatcgcg tatttcgcct cgctcaggcg caatcacgaa tgaataacgg
     5761 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     5821 gaaagaaatg cataaactgt tgccattctc accggattca gtcgtcactc atggtgattt
     5881 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     5941 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     6001 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     6061 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     6121 gtaacactgg cgagctctgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6181 ctataaaaat aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc
     6241 taaggatgat ttctgcacct gcaccagtca gtaaaacgac ggccagtagt caaaagcctc
     6301 cgaccggagg cttttgactt ggttcaggtg gagtgggag
//
