LOCUS       sequence-464164-        5912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of cyanovirin-N homolog (CV-N 
            homolog) in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5912)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 5912)
  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_3HNX
FEATURES             Location/Qualifiers
     source          1..5912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(590..619)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             642..664
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    679..703
                     /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(703..722)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(704..722)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(810..829)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1035..1112
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1113..2195
                     /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(1132..1151)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3004..3195
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3189..3211)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3297..3436
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3352..3371
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3451..3468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3622..4210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3702..3721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4321..4340)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4332..5147
                     /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             4332..5147
                     /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(4405..4424)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5239..5694)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5376..5397)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5588..5607
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(5731..5778)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     5774..5792
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(5845..5862)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gattcaggtt aacatcacga tcgtggaatt cgccattaca gtcgcgtaaa cgggctctca
       61 gaatcggctg ctccgccgca ccttccttcg ggtgaaaacg gatgtcctct gcggtttccg
      121 tgaagttctg accaccccat tgaaaatgac catcgttgtt gccgatgata gtgtccaggt
      181 ccagttcgct ggtcacccag ttgccatccg cattacggca ctcagcgcgc agggtacgac
      241 cgccattggt caatactgca ttcctagaac tatcagcata tgatccagag ccgccaatct
      301 gttcacggtg ggcttcaatg atgtcgttgt cttccatgtc caggtcttcc ggcgcttgat
      361 ctgcctggat gcgaataccg tcatacagga aacgcaggct atccatttcc ttgccctgac
      421 gtttggcaaa cgcttccatc aggcgacgca gcggcgtggt ctttttgatc ttaaagaaaa
      481 tttcagagct gccgtcacta actttcaggt taatatgggt ttccggtttc acttccggct
      541 tgacttccgg ctttgcttct tggttgactt ccgaatcttg caggctaccg tggtgatggt
      601 gatggtgatg atggtgatga atcttcgcca tcatatgtat atctccttct taaagttaaa
      661 caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat
      721 taatttcgcg ggatcgagat cgatctcgat cctctacgcc ggacgcatcg tggccggcat
      781 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
      841 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
      901 cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt
      961 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
     1021 gcgtcgagat cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc
     1081 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
     1141 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     1201 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     1261 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     1321 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     1381 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     1441 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     1501 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     1561 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     1621 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     1681 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     1741 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
     1801 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
     1861 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gacatctcgg
     1921 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     1981 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     2041 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
     2101 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     2161 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
     2221 ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc
     2281 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     2341 atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt
     2401 cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc
     2461 gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc
     2521 gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta
     2581 ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga
     2641 agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc
     2701 cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc
     2761 tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc
     2821 tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta
     2881 ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc
     2941 ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca
     3001 tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca
     3061 ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa
     3121 tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac
     3181 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     3241 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     3301 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag
     3361 agcagattgt actgagagtg caccattgcg gtgtgaaata ccgcacagat gcgtaaggag
     3421 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3481 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3541 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3601 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3661 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3721 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3781 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3841 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3901 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3961 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4021 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     4081 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4141 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4201 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4261 ctcacgttaa gggattttgg tcatgaacaa taaaactgtc tgcttacata aacagtaata
     4321 caaggggtgt tatgagccat attcaacggg aaacgtcttg ctctaggccg cgattaaatt
     4381 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     4441 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     4501 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     4561 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     4621 tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag
     4681 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     4741 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     4801 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     4861 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     4921 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     4981 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     5041 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     5101 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaagaa ttaattcatg
     5161 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5221 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     5281 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     5341 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     5401 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     5461 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     5521 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     5581 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     5641 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     5701 cgccaatccg gatatagttc ctcctttcag caaaaaaccc ctcaagaccc gtttagaggc
     5761 cccaaggggt tatgctagtt attgctcagc ggtggcagca gccaactcag cttcctttcg
     5821 ggctttgtta gcagccggat ctcagtggtg gtggtggtgg tgctcgagtc attacgaacc
     5881 gaaaaccaag cggccgttca cgttttgaat ac
//
