LOCUS       sequence-70448-e        6417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pET29a_PPP CODA PFV Integrase with C-terminal His tag (GPG removed 
            from pLZL2373).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6417)
  AUTHORS   Qi X, Vargas E, Larsen L, Knapp W, Hatfield GW, Lathrop R, Sandmeyer
            S
  TITLE     Directed DNA shuffling of retrovirus and retrotransposon integrase 
            protein domains.
  JOURNAL   PLoS One. 2013 May 17;8(5):e63957. doi: 
            10.1371/journal.pone.0063957. Print 2013.
  PUBMED    23691126
REFERENCE   2  (bases 1 to 6417)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2013-05-17"; volume: "8(5):e63957. doi"; pages: " 
            10.1371/journal.pone.0063957. Print 2013"
COMMENT     Sequence Label: pLZL2386
FEATURES             Location/Qualifiers
     source          1..6417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..24
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(77..95)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      91..138
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      175..630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     472..493
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(723..1538)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(723..1538)
                     /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     1446..1465
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1530..1549
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1660..2248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2149..2168
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2402..2419
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2434..2576
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2502..2521)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2662..2684
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2678..2869)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3678..4760)
                     /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"
     primer_bind     4722..4741
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4761..4838)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5044..5063
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5151..5170
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5151..5169
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5170..5194
                     /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)."
     RBS             5209..5231
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
       61 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
      121 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
      181 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
      241 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
      301 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
      361 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
      421 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
      481 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
      541 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
      601 attttaacaa aatattaacg tttacaattt caggtggcac ttttcgggga aatgtgcgcg
      661 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
      721 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
      781 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
      841 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
      901 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
      961 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     1021 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1081 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     1141 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1201 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     1261 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     1321 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     1381 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     1441 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     1501 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     1561 atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca
     1621 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1681 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1741 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1801 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1861 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1921 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1981 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2041 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2101 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2161 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2221 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2281 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2341 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2401 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2461 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2521 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2581 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2641 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2701 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2761 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2821 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2881 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2941 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     3001 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     3061 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     3121 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     3181 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     3241 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     3301 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
     3361 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     3421 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     3481 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     3541 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     3601 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     3661 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3721 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     3781 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     3841 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     3901 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     3961 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     4021 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     4081 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     4141 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     4201 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     4261 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     4321 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     4381 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     4441 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     4501 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     4561 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     4621 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     4681 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     4741 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     4801 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     4861 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     4921 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     4981 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     5041 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     5101 cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact
     5161 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     5221 taagaaggag atatacatat gtgcaacact aaaaagccaa acctggatgc tgaattggat
     5281 caactgcttc agggccacta cattaaaggt tacccaaaac agtacaccta cttcctggaa
     5341 gacggtaaag taaaagtttc tcgtccagaa ggtgttaaaa ttattccacc acagtctgat
     5401 cgccagaaaa ttgttttgca ggctcacaac ctggctcaca ctggtcgtga ggctactctg
     5461 ctgaaaattg ctaacctgta ctggtggcca aacatgcgta aggatgttgt taaacagttg
     5521 ggtcgttgcc agcaatgcct gattactaac gcatctaaca aagcatctgg tccaattttg
     5581 cgtccggatc gtccacagaa accattcgat aaattcttca ttgattacat tggtccgctg
     5641 cctccatctc agggttacct gtacgttttg gttgtagttg atggtatgac cggtttcact
     5701 tggctgtacc caactaaagc tccatccact tctgcaactg ttaagtcctt gaacgttctg
     5761 acctccattg ctattccaaa agttattcac tctgatcaag gcgctgcttt tacttcttcc
     5821 actttcgctg aatgggctaa agaacgtggt attcaccttg aattttccac cccatatcac
     5881 ccgcagtctt ctggtaaagt tgaacgtaaa aactccgaca ttaaacgtct gctgactaag
     5941 ctgctggtag gtcgtccgac taaatggtac gatctgttac cggttgttca actggctctg
     6001 aacaacactt actctccagt actgaaatac actccacacc agctcctgtt cggtattgac
     6061 tctaacactc ctttcgctaa ccaggatact ctggatctga ctcgtgaaga agaactgtcc
     6121 ttgttgcagg aaattcgtac ctctctgtac cacccgtcta ctccgccagc ttcttctcgt
     6181 tcctggagcc cagttgttgg tcagttggtt caggaacgtg ttgctcgtcc tgcttctctg
     6241 cgccctcgtt ggcacaaacc atctactgtt ctgaaagttt tgaacccacg caccgttgtt
     6301 attctggatc atctgggtaa caaccgtact gtttccattg ataacctgaa accaacctct
     6361 caccagaacg gtactactaa cgatactgct actatggatc acttggaaaa gaacgaa
//
