LOCUS       sequence-464130-        6122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of pre-mRNA-splicing factor 38A in 
            Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6122)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 6122)
  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_4RZ9
FEATURES             Location/Qualifiers
     source          1..6122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             29..46
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(99..117)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      113..160
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      197..652
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(284..303)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     494..515
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(744..1559)
                     /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(744..1559)
                     /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     1467..1486
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1551..1570
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1681..2269
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2170..2189
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2423..2440
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2455..2594
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2520..2539)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2680..2702
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2696..2887)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3696..4778)
                     /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     4740..4759
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4779..4856)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5062..5081
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5169..5188
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5169..5187
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5188..5212
                     /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             5227..5249
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5272..5301
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
ORIGIN
        1 ggagcagctg ggttcgtaat gactcgagca ccaccaccac caccactgag atccggctgc
       61 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
      121 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
      181 cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
      241 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
      301 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
      361 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
      421 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
      481 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
      541 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
      601 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
      661 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      721 tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat
      781 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
      841 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
      901 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
      961 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     1021 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     1081 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     1141 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     1201 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     1261 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     1321 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     1381 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     1441 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     1501 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata
     1561 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc
     1621 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1681 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1741 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1801 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     1861 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1921 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1981 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2041 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2101 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2161 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2221 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2281 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2341 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2401 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     2461 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     2521 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     2581 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2641 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     2701 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     2761 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     2821 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     2881 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     2941 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     3001 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     3061 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     3121 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     3181 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     3241 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     3301 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg
     3361 ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga
     3421 aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg
     3481 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
     3541 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
     3601 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     3661 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3721 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3781 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     3841 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     3901 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     3961 tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     4021 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     4081 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     4141 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     4201 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     4261 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     4321 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     4381 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     4441 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     4501 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     4561 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     4621 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     4681 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     4741 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     4801 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     4861 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     4921 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     4981 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     5041 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     5101 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc
     5161 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
     5221 aataattttg tttaacttta agaaggagat atacatatga tggcgaagat tcatcaccat
     5281 catcaccatc accatcacca cggtagcctg caagattcgg aagtcaacca agaagcaaag
     5341 ccggaagtca agccggaagt gaaaccggaa acccatatta acctgaaagt tagtgacggc
     5401 agctctgaaa ttttctttaa gatcaaaaag accacgccgc tgcgtcgcct gatggaagcg
     5461 tttgccaaac gtcagggcaa ggaaatggat agcctgcgtt tcctgtatga cggtattcgc
     5521 atccaggcag atcaagcgcc ggaagacctg gacatggaag acaacgacat cattgaagcc
     5581 caccgtgaac agattggcgg ctctggaaat aggacagtaa aagatgctca ctcaatacac
     5641 ggcaccaacc cgcagtattt ggtagaaaag atcattcgta cccgcattta tgaaagcaaa
     5701 tactggaaag aagagtgctt cggcctgacg gccgagttag tcgtggacaa agcaatggaa
     5761 ctgcgttttg ttggtggtgt ttatggcggt aatattaagc cgaccccatt tctgtgcctg
     5821 accctgaaaa tgttgcaaat tcagccggaa aaagatatca tcgtggagtt cattaagaat
     5881 gaagatttta agtacgttcg tatgctgggt gctctgtaca tgagactgac cggtactgcg
     5941 atcgactgtt ataaatactt ggagccgttg tacaacgatt accgcaagat caagtcccaa
     6001 aaccgcaacg gcgaattcga gctgatgcat gtggacgagt tcatcgacga attgcttcac
     6061 agcgaacgtg tttgcgatat tatcctgccg cgtctccaaa aacgttatgt gctggaggag
     6121 gc
//
