LOCUS       sequence-464127-        5957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of paired immunoglobulin-like type 2
            receptor alpha in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5957)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 5957)
  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_4NFB
FEATURES             Location/Qualifiers
     source          1..5957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(14..829)
                     /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(14..829)
                     /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     737..756
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     821..840
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      951..1539
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1440..1459
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1693..1710
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1725..1864
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1790..1809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1950..1972
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1966..2157)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2966..4048)
                     /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     4010..4029
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4049..4126)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4332..4351
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4439..4458
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4439..4457
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4458..4482
                     /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             4497..4519
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4542..4571
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5256..5273
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5326..5344)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5340..5387
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5424..5879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5511..5530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5721..5742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
       61 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
      121 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
      181 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
      241 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
      301 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
      361 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
      421 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
      481 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
      541 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
      601 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
      661 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
      721 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
      781 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg
      841 tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag
      901 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
      961 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1021 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1081 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     1141 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1201 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1261 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1321 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1381 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1441 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1501 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1561 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1621 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1681 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     1741 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
     1801 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     1861 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     1921 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     1981 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     2041 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     2101 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     2161 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     2221 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     2281 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     2341 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     2401 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     2461 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     2521 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     2581 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc
     2641 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
     2701 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
     2761 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     2821 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
     2881 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     2941 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3001 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     3061 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     3121 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     3181 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     3241 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     3301 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     3361 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     3421 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     3481 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     3541 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     3601 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     3661 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     3721 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     3781 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     3841 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     3901 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     3961 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     4021 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     4081 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     4141 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     4201 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     4261 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     4321 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     4381 atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc cgcgaaatta
     4441 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg
     4501 tttaacttta agaaggagat atacatatga tggcgaagat tcatcaccat catcaccatc
     4561 accatcacca cggtagcctg caagattcgg aagtcaacca agaagcaaag ccggaagtca
     4621 agccggaagt gaaaccggaa acccatatta acctgaaagt tagtgacggc agctctgaaa
     4681 ttttctttaa gatcaaaaag accacgccgc tgcgtcgcct gatggaagcg tttgccaaac
     4741 gtcagggcaa ggaaatggat agcctgcgtt tcctgtatga cggtattcgc atccaggcag
     4801 atcaagcgcc ggaagacctg gacatggaag acaacgacat cattgaagcc caccgtgaac
     4861 agattggcgg ctctggaatg ctatatggag taacacaacc caaacacttg tctgctagca
     4921 tgggtggttc cgtggagatc ccgtttagct tctactaccc gtgggagctg gcgaccgcac
     4981 cggatgttcg tatctcctgg cgtcgtggtc atttccaccg ccagtcgttc tatagcaccc
     5041 gtccgccatc gatccacaag gactacgtga accgcctgtt tctcaactgg accgaaggcc
     5101 agaaaagcgg ttttctgcgt attagcaatc tgcagaagca agatcagtct gtttatttct
     5161 gccgcgtcga gttggacact agaagcagcg gccgtcaaca atggcagtca attgaaggca
     5221 cgaaactgtc cattaccggt tcgtaatgac tcgagcacca ccaccaccac cactgagatc
     5281 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     5341 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     5401 ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5461 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5521 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5581 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5641 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5701 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5761 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5821 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     5881 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5941 tcaaatatgt atccgct
//
