LOCUS       sequence-464122-        5990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of fatty acid-binding protein, 
            adipocyte (ALBP) in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5990)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 5990)
  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_3RZY
FEATURES             Location/Qualifiers
     source          1..5990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..21
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(145..192)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     188..206
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(259..276)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(994..1023)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             1046..1068
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1083..1107
                     /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(1107..1126)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1108..1126)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1214..1233)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1439..1516
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1517..2599
                     /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(1536..1555)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3408..3599
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3593..3615)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3701..3840
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3756..3775
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3855..3872)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4026..4614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4106..4125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4725..4744)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4736..5551
                     /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             4736..5551
                     /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(4809..4828)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(join(5643..5990,1..108))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5780..5801)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
       61 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccaa
      121 tccggatata gttcctcctt tcagcaaaaa acccctcaag acccgtttag aggccccaag
      181 gggttatgct agttattgct cagcggtggc agcagccaac tcagcttcct ttcgggcttt
      241 gttagcagcc ggatctcagt ggtggtggtg gtggtgctcg agtcattacg aacccgcacg
      301 ttcataaaca cgcgtgctgg ttacaccctt cataacgcac tcaacaacca atttgtcatc
      361 ttcgcgcttg cgcttaatgg ttgtagattt gccatcccat ttctgcacgt gcaccagcac
      421 accgccgtcc agggtgatcg tggatttcac cttacgatca tcagcggtca cctcgtcgaa
      481 ctcttgaccc aggataaagg aaatttcggt gtttttaaag gtgctctccg acttgatagt
      541 aatcacatca ccattaacgc tgataatcat gttcggtttt gccatacccg cgaccttacg
      601 ggtggcgaag cccacgccaa cttctttcat gtagtcgtcg aagttctcgc tgctgactag
      661 tttccatgtt cctacaaaag catcacatcc agagccgcca atctgttcac ggtgggcttc
      721 aatgatgtcg ttgtcttcca tgtccaggtc ttccggcgct tgatctgcct ggatgcgaat
      781 accgtcatac aggaaacgca ggctatccat ttccttgccc tgacgtttgg caaacgcttc
      841 catcaggcga cgcagcggcg tggtcttttt gatcttaaag aaaatttcag agctgccgtc
      901 actaactttc aggttaatat gggtttccgg tttcacttcc ggcttgactt ccggctttgc
      961 ttcttggttg acttccgaat cttgcaggct accgtggtga tggtgatggt gatgatggtg
     1021 atgaatcttc gccatcatat gtatatctcc ttcttaaagt taaacaaaat tatttctaga
     1081 ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg
     1141 agatcgatct cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     1201 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
     1261 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
     1321 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
     1381 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga
     1441 caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     1501 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     1561 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     1621 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     1681 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     1741 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     1801 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     1861 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     1921 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     1981 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     2041 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     2101 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     2161 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     2221 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     2281 cgccattacc gagtccgggc tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga
     2341 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     2401 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     2461 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
     2521 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     2581 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     2641 cgggatctcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     2701 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     2761 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     2821 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     2881 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccc
     2941 cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc
     3001 ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca
     3061 aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc
     3121 tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct
     3181 gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag
     3241 tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca
     3301 gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg
     3361 gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg
     3421 aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga
     3481 aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg
     3541 ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     3601 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     3661 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     3721 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     3781 agtgcaccat tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     3841 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3901 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3961 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4021 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4081 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4141 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4201 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4261 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4321 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4381 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4441 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4501 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4561 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4621 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4681 ttggtcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     4741 ccatattcaa cgggaaacgt cttgctctag gccgcgatta aattccaaca tggatgctga
     4801 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     4861 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     4921 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     4981 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     5041 cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     5101 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     5161 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     5221 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     5281 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     5341 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     5401 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     5461 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     5521 gtttcatttg atgctcgatg agtttttcta agaattaatt catgagcgga tacatatttg
     5581 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5641 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     5701 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     5761 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5821 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5881 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5941 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
//
