LOCUS       Exported                5715 bp ds-DNA     circular SYN 17-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5715)
  AUTHORS   Manuel Iburg
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 18, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5715
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      13..468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(561..1376)
                     /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"
     primer_bind     complement(1473..1506)
                     /label=Vector.REV
     primer_bind     1491..1517
                     /label=Fragment.FOR
     rep_origin      1498..1505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      1507..2043
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2028..2060)
                     /label=Fragment.REV
     primer_bind     2036..2070
                     /label=Vector.FOR
     rep_origin      2048..2055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2241..2380
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2482..2673)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3482..4564)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4565..4642)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(4565..4581)
                     /label=Simluate FKBP F
     promoter        4951..4969
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4970..4994
                     /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     complement(5001..5038)
                     /label=pET26-Gib2-R
                     /note="Ta = 62.5Tm = 61.5"
     primer_bind     complement(5001..5038)
                     /label=pET26b-OptHis-Gib-R
                     /note="Ta = 63.4Tm = 60.4"
     primer_bind     complement(5012..5067)
                     /label=p26-5HisMoLinker-R
                     /note="Ta = 60.4Tm = 57.4"
     primer_bind     complement(5018..5042)
                     /label=MI003 SmNterm rev
     primer_bind     5019..5057
                     /label=pET-FA3-IF
                     /note="Ta = 64.9Tm = 63.9"
     CDS             5051..5068
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5072..5104
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             5108..5131
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(5120..5140)
                     /label=Primer 1
     primer_bind     complement(5120..5139)
                     /label=pBAD-ddXFP-VR
                     /note="Ta = 62.6Tm = 61.6"
     primer_bind     5121..5158
                     /label=pBAD-NP-IF
                     /note="Ta = 64.5Tm = 66.8"
     primer_bind     complement(5123..5140)
                     /label=ddXFP-TSLZ-VR B
                     /note="Ta= 61.3Tm = 64.7"
     primer_bind     complement(5126..5140)
                     /label=pBAD-T-ddXFPB-VR
                     /note="Ta = 65.2Tm = 64.2"
     primer_bind     5139..5149
                     /label=NLNP-SwapP-VF
                     /note="Ta = 68.5Tm = 69.3"
     primer_bind     5139..5149
                     /label=Simluate FKBP F
     CDS             5141..5461
                     /codon_start=1
                     /label=FKBP12
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     primer_bind     5141..5158
                     /label=Gibson060
     primer_bind     5141..5158
                     /label=pTTGFP-FKBP-F2F
                     /note="Ta = 63.1Tm = 64.6"
     misc_difference 5209
                     /label=mut to T
     primer_bind     complement(5443..5461)
                     /label=pTTGFP-FKBP-F2R
                     /note="Ta = 63.1Tm = 60.1"
     primer_bind     complement(5444..5505)
                     /label=pLQ-FKBP-NLNP-R
                     /note="Ta = 60Tm = 56"
     primer_bind     complement(5444..5461)
                     /label=pLQ-FKBP-ddR-VR
                     /note="Ta = 60Tm = 57"
     primer_bind     5462..5480
                     /label=MI003 SmNterm for
     primer_bind     5462..5480
                     /label=Gibson039
     primer_bind     complement(5490..5511)
                     /label=Fix-NLNP-R
                     /note="Ta = 69Tm = 66"
     primer_bind     complement(5498..5515)
                     /label=Gibson050
     primer_bind     5506..5568
                     /label=Gibson049
     primer_bind     5512..5529
                     /label=Fix-NLNP-F
                     /note="Ta = 69Tm = 67"
     CDS             5516..5548
                     /codon_start=1
                     /label=NanoBiT Small Fragment
                     /translation="VTGYRLFEEIL"
     primer_bind     5516..5533
                     /label=Gibson074
     primer_bind     complement(5516..5532)
                     /label=MI003 SmNterm rev
     primer_bind     complement(5531..5597)
                     /label=Gibson041
     primer_bind     complement(5531..5571)
                     /label=pBAD-NP-IR
                     /note="Ta = 64.5Tm = 63.5"
     primer_bind     5533..5548
                     /label=MI003 SmNterm for
     primer_bind     5552..5569
                     /label=pBAD-ddXFP-VF
                     /note="Ta = 62.6Tm = 61.6"
     primer_bind     complement(5556..5578)
                     /label=rrnB-F1060_Seq-R
     primer_bind     complement(5558..5597)
                     /label=pBAD2pETv2
                     /note="Ta = 64Tm = 63"
     primer_bind     5578..5599
                     /label=pET26b-ChangeHis-F
                     /note="Ta = 68Tm = 65"
     primer_bind     5578..5595
                     /label=pET26-Gib2-F
                     /note="Ta = 62.5Tm = 64"
     primer_bind     5578..5595
                     /label=p26-5HisMoLinker-F
                     /note="Ta = 60.4Tm = 62"
     terminator      5644..5691
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
       61 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
      121 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
      181 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
      241 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
      301 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
      361 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
      421 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttca ggtggcactt
      481 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
      541 atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
      601 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
      661 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
      721 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
      781 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
      841 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
      901 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
      961 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     1021 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     1081 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     1141 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     1201 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     1261 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     1321 atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca
     1381 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta
     1441 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1501 agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg
     1561 cggtttttcg aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg
     1621 agcgcagtca ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac
     1681 taactcctct aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg
     1741 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt
     1801 cgtgcataca gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa
     1861 tgagacaaac gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga
     1921 gagcgcacga gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1981 cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg
     2041 aaaaacgtat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2101 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2161 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     2221 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2281 tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     2341 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     2401 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2461 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     2521 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     2581 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     2641 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     2701 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     2761 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     2821 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     2881 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     2941 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     3001 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     3061 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     3121 cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa
     3181 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
     3241 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
     3301 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     3361 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     3421 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     3481 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3541 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     3601 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     3661 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     3721 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     3781 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     3841 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     3901 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     3961 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     4021 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     4081 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     4141 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     4201 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     4261 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     4321 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     4381 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     4441 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     4501 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     4561 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     4621 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     4681 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     4741 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     4801 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     4861 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     4921 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
     4981 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat
     5041 ggggggttct catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat
     5101 gggtcgggat ctgtacgacg atgacgataa ggatccgagc ggtgtgcagg tggaaaccat
     5161 tagcccgggt gatggccgta cctttccgaa acgtggccag acctgcgtgg tgcattatac
     5221 cggcatgctg gaagatggca aaaaatttga tagcagccgc gatcgtaaca aaccgttcaa
     5281 attcatgctg ggcaaacagg aagtgattcg cggctgggag gaaggcgtgg cgcagatgag
     5341 cgttggccag cgtgcgaaac tgaccatcag cccggattat gcgtatggcg cgaccggcca
     5401 tccgggtatt attccgccgc atgcgaccct ggtgtttgat gtggaactgc tgaaactgga
     5461 agtttctcaa ggcagttcag gtggtggcgg gagcggaggt ggaggctcga gcggtgtgac
     5521 cggctaccgg ctgttcgagg agattctgtg aggctgtttt ggcggatgag agaagattga
     5581 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     5641 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     5701 ggaactatat ccgga
//
