LOCUS       sequence-464116-        6539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of malonyl CoA-acyl carrier protein 
            transacylase in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6539)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 6539)
  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_5YPV
FEATURES             Location/Qualifiers
     source          1..6539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      41..496
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(128..147)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     338..359
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(588..1403)
                     /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(588..1403)
                     /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     1311..1330
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1395..1414
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1525..2113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2014..2033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2267..2284
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2299..2438
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2364..2383)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2524..2546
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2540..2731)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3540..4622)
                     /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     4584..4603
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4623..4700)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4906..4925
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5013..5032
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5013..5031
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5032..5056
                     /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             5071..5093
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5116..5145
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             6412..6429
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6482..6500)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      join(6496..6539,1..4)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
       61 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
      121 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
      181 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
      241 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
      301 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
      361 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      421 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      481 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      541 tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca
      601 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
      661 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
      721 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
      781 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
      841 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
      901 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
      961 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     1021 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     1081 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     1141 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     1201 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     1261 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     1321 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     1381 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     1441 tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1501 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1561 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1621 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1681 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1741 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1801 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1861 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1921 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1981 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2041 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2101 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2161 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2221 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2281 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2341 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     2401 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     2461 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2521 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     2581 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     2641 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     2701 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     2761 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     2821 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     2881 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     2941 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     3001 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     3061 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     3121 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     3181 cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     3241 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     3301 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     3361 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     3421 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     3481 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     3541 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3601 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     3661 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     3721 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     3781 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     3841 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     3901 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     3961 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     4021 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     4081 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     4141 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     4201 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     4261 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     4321 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     4381 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     4441 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     4501 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     4561 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     4621 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     4681 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     4741 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     4801 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     4861 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     4921 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     4981 gaggatcgag atcgatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
     5041 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat
     5101 atgatggcga agattcatca ccatcatcac catcaccatc accacggtag cctgcaagat
     5161 tcggaagtca accaagaagc aaagccggaa gtcaagccgg aagtgaaacc ggaaacccat
     5221 attaacctga aagttagtga cggcagctct gaaattttct ttaagatcaa aaagaccacg
     5281 ccgctgcgtc gcctgatgga agcgtttgcc aaacgtcagg gcaaggaaat ggatagcctg
     5341 cgtttcctgt atgacggtat tcgcatccag gcagatcaag cgccggaaga cctggacatg
     5401 gaagacaacg acatcattga agcccaccgt gaacagattg gcggctctgg agcggcaaca
     5461 aaaactgctt tcgtatttcc cggccaaggt tcccaaaaag tgggcatgct ggcggagctg
     5521 gcggagcaat tttctggtgt aggtcagacc ttcgcggaag caagcgacgc gttgggcttc
     5581 gacctgtggc acattgcgca atccggcgaa ggtctggacc agacggaaaa cacccagccg
     5641 gtgttgctga cggcatcgat cgcgttgtgg cgtctgtggc tggacctggg cggtttggct
     5701 ccgaaatacc tggcgggtca tagtctgggt gaatatagcg cattggtcgc ggcgggtagc
     5761 atgagcctag cggacgccgt gaagctcgtg aacctgcgtg gtaaactgat gcaaaacgcg
     5821 gttccgcagg gcgagggtgc tatggcagcc atcctggggc tggcggacga caaggtggtt
     5881 gagctgtgtc agtctgtgag cgcactcggt caaggctcgg tcgaggcggc gaattataac
     5941 agccagggcc aggttgttat cgctggctcc accgcatcag tgcagcaggt tatggcttta
     6001 gcgaaggaac agtccgcgaa ggcgatcgct ctaccggtta gcgtgccgag ccactgcagc
     6061 ctgatgaaac cagcagcgga gaagttcgcc gaggcgctgg aacaaacggc tatcgaattg
     6121 ccgaccattc cggtcatcca aaacgttaat gcaggtatcg cgaccgatgt tgcacaactt
     6181 cgccaggcgt tgactgccca attataccag tctgtgcagt ggacccgcac catgcaatcc
     6241 ctgcaggatc aaggtattca gtacattgtc gagtgcggtc cgggcaccgt tctgagcaat
     6301 ctggcaaaac gtctgccaaa tattgaaaag gccttttcta ttgatagcaa aagcaagatg
     6361 gaggatgcgc tgaacgccgt tctggtggcc gcgggttcgt aatgactcga gcaccaccac
     6421 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     6481 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggttt
//
