LOCUS       sequence-464131-        6065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of cell division control protein 73 
            in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6065)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 6065)
  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"
COMMENT     Sequence Label: pAIP4_3V46
FEATURES             Location/Qualifiers
     source          1..6065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    65..204
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     120..139
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(219..236)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(390..978)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(470..489)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1089..1108)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1100..1915
                     /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             1100..1915
                     /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(1173..1192)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2007..2462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2144..2165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2356..2375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(2499..2546)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2542..2560
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2613..2630)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3423..3452)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             3475..3497
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3512..3536
                     /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(3536..3555)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3537..3555)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3643..3662)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3868..3945
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3946..5028
                     /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     complement(3965..3984)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5837..6028
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6022..6044)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
       61 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
      121 ggcatcagag cagattgtac tgagagtgca ccattgcggt gtgaaatacc gcacagatgc
      181 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
      241 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
      301 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
      361 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      421 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      481 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      541 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      601 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
      661 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
      721 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
      781 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
      841 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
      901 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
      961 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1021 aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg cttacataaa
     1081 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg
     1141 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     1201 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     1261 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     1321 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     1381 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     1441 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     1501 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     1561 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     1621 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     1681 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     1741 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     1801 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     1861 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagaatt
     1921 aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1981 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     2041 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2101 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2161 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2221 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2281 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2341 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2401 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2461 gtcccattcg ccaatccgga tatagttcct cctttcagca aaaaacccct caagacccgt
     2521 ttagaggccc caaggggtta tgctagttat tgctcagcgg tggcagcagc caactcagct
     2581 tcctttcggg ctttgttagc agccggatct cagtggtggt ggtggtggtg ctcgagtcat
     2641 tacgaaccgt agccacggct gatcaactct ttctccaggc tatgccagaa ataacgaacc
     2701 acctctacgt ccttgaagcg tttgtttttg tcgagttcca ccttctcaac gttccactgt
     2761 tgcacgtgtt gcggcacgga gtcaccagca aagtggaagt agtaaccttt gcagcgctga
     2821 aacaattctt gcggggaatt ccactgatag ttgttgaact gccaggtatg accagtcgta
     2881 aaaattgcca ccacacgatc ccagtattcc ggcttggtga acatacgggt attatcaacg
     2941 atgataaaac gaatagggcg gctaattctt tcaaagttct tttcaatgtt aaccaggcca
     3001 tttggaacgc tcggcagatt acgcgggttg acatacttgc tctccagcag gaactgtttg
     3061 atgttcgcaa cggtcaggat cgaagaagct gcacttggta ttagaattat gggatctcca
     3121 gagccgccaa tctgttcacg gtgggcttca atgatgtcgt tgtcttccat gtccaggtct
     3181 tccggcgctt gatctgcctg gatgcgaata ccgtcataca ggaaacgcag gctatccatt
     3241 tccttgccct gacgtttggc aaacgcttcc atcaggcgac gcagcggcgt ggtctttttg
     3301 atcttaaaga aaatttcaga gctgccgtca ctaactttca ggttaatatg ggtttccggt
     3361 ttcacttccg gcttgacttc cggctttgct tcttggttga cttccgaatc ttgcaggcta
     3421 ccgtggtgat ggtgatggtg atgatggtga tgaatcttcg ccatcatatg tatatctcct
     3481 tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta
     3541 tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac gccggacgca
     3601 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
     3661 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
     3721 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
     3781 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
     3841 cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt
     3901 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     3961 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     4021 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     4081 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     4141 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     4201 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     4261 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     4321 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     4381 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     4441 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     4501 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     4561 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     4621 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     4681 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     4741 gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     4801 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     4861 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     4921 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     4981 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     5041 taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt
     5101 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     5161 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg
     5221 cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat
     5281 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa
     5341 gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc tcctgtcgtt
     5401 gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg
     5461 cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat
     5521 ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat
     5581 tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt
     5641 attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac
     5701 cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg
     5761 tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc
     5821 ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat
     5881 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc
     5941 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg
     6001 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     6061 tctgt
//
