LOCUS       sequence-464197-        6035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of ADP-ribose glycohydrolase OARD1 
            in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6035)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 6035)
  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_4J5Q
FEATURES             Location/Qualifiers
     source          1..6035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..72)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     263..282
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(406..453)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     449..467
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(520..537)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1300..1329)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             1352..1374
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1389..1413
                     /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(1413..1432)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1414..1432)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1520..1539)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1745..1822
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1823..2905
                     /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(1842..1861)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3714..3905
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3899..3921)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4007..4146
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4062..4081
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4161..4178)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4332..4920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4412..4431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5031..5050)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5042..5857
                     /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             5042..5857
                     /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(5115..5134)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(join(5949..6035,1..369))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
       61 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
      121 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
      181 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
      241 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
      301 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
      361 agggcgcgtc ccattcgcca atccggatat agttcctcct ttcagcaaaa aacccctcaa
      421 gacccgttta gaggccccaa ggggttatgc tagttattgc tcagcggtgg cagcagccaa
      481 ctcagcttcc tttcgggctt tgttagcagc cggatctcag tggtggtggt ggtggtgctc
      541 gagtcattac gaacccaggg tataaacggt gatcttgatg tcggtcgcct caaaaacttc
      601 ctcaatcata gcgctcacgt tttcccactg gagacgatcc agaccgcagc caatacgcgg
      661 catagataag tcagtaacgc cgttcttcag gcagtgggat ttcatggctt ccagcgactt
      721 ttgcaggttc tcgtacgtcg gtttgtggct agcgcgcttt ttggtaatca ggtagtagat
      781 ataacggccg tcgcgtttaa gcaccgcaac ctcaccggac ttcttttgct gattcaacag
      841 ctcctgcaca ccgccgaact tctttttaaa caacaccgca atacctgcgc ccatacgaca
      901 atcttcgctg atgcaatgtg ccaggctgtc ggttttcgga cacgcgaaca aatcaccttt
      961 gacatatgtt atccttgatc cttcgggatc ctctccagag ccgccaatct gttcacggtg
     1021 ggcttcaatg atgtcgttgt cttccatgtc caggtcttcc ggcgcttgat ctgcctggat
     1081 gcgaataccg tcatacagga aacgcaggct atccatttcc ttgccctgac gtttggcaaa
     1141 cgcttccatc aggcgacgca gcggcgtggt ctttttgatc ttaaagaaaa tttcagagct
     1201 gccgtcacta actttcaggt taatatgggt ttccggtttc acttccggct tgacttccgg
     1261 ctttgcttct tggttgactt ccgaatcttg caggctaccg tggtgatggt gatggtgatg
     1321 atggtgatga atcttcgcca tcatatgtat atctccttct taaagttaaa caaaattatt
     1381 tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg
     1441 ggatcgagat cgatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
     1501 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
     1561 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
     1621 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
     1681 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat
     1741 cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     1801 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     1861 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     1921 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     1981 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     2041 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     2101 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     2161 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     2221 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     2281 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     2341 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     2401 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     2461 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     2521 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     2581 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata
     2641 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
     2701 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     2761 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     2821 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2881 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt
     2941 aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     3001 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     3061 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     3121 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     3181 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     3241 cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg
     3301 gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct
     3361 gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt
     3421 aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag
     3481 gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac
     3541 cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac
     3601 gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt
     3661 tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca
     3721 aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc
     3781 tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg
     3841 accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     3901 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     3961 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     4021 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     4081 actgagagtg caccattgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     4141 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4201 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4261 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4321 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4381 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4441 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4501 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4561 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4621 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4681 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4741 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     4801 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4861 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4921 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4981 gggattttgg tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     5041 tatgagccat attcaacggg aaacgtcttg ctctaggccg cgattaaatt ccaacatgga
     5101 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     5161 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     5221 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     5281 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     5341 gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     5401 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     5461 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     5521 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     5581 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     5641 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     5701 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     5761 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     5821 attgcagttt catttgatgc tcgatgagtt tttctaagaa ttaattcatg agcggataca
     5881 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5941 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     6001 cagctcattt tttaaccaat aggccgaaat cggca
//
