LOCUS       sequence-7395-ex        6510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6510)
  AUTHORS   Ha JY, Lee JH, Kim KH, Kim DJ, Lee HH, Kim HK, Yoon HJ, Suh SW
  TITLE     Overexpression, crystallization and preliminary X-ray 
            crystallographic analysis of erythronate-4-phosphate dehydrogenase 
            from Pseudomonas aeruginosa.
  JOURNAL   Acta Crystallograph Sect F Struct Biol Cryst Commun. 2006 Feb 1. 
            62(Pt 2):139-41.
  PUBMED    16511285
REFERENCE   2  (bases 1 to 6510)
  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"; journalName: "Acta 
            Crystallograph Sect F Struct Biol Cryst Commun. 2006 Feb 1. 62(Pt 
            2):139-41."
COMMENT     Sequence Label: pET21a(+)-PA PdxB
FEATURES             Location/Qualifiers
     source          1..6510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1314..1336
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1351..1375
                     /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(1375..1394)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1376..1394)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1478..1497)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1703..1780
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1781..2863
                     /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(1800..1819)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3672..3863
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3857..3879)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3965..4107
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4020..4039
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4122..4139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4293..4881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4373..4392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5052..5843,5844..5912))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5675..5694
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5913..6017)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6044..6499)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6181..6202)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6393..6412
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagctagacc aactgcgccc
      181 ccagcgcccg caccacccgc tgcaactccg cctgtccacc ggtggcgacg cgcaggccgg
      241 tgatctcgcg gcgcggcggg tagtgtttgc gcagggcgtc gaaggcggcg cggcgggtgg
      301 cgctgtcgcc ggtcaggctg cggcggaagg cggcgtcgtc gctgcgcggg tcgtacaccg
      361 cgcggcacag cgtggccagc gcccaggccg ggtcgcagcc ggggttgagt tgcaggccgg
      421 cgagccaggt ttccggcagc acgtcttgca ggctcacgcg ttcggcgatg ccgcgccagg
      481 cgcagtaggc ctggtagatc tgtgcggtgc cgcgcagctt gccttccagg ctgtagccgg
      541 cgatgtgcgg cgtggcgatc aggcagcgcg cggccagttc ggggtcggcc tgcggttcgc
      601 cttcccagac gtccagcgcc acttccaggt cggcgccgcc ttcgagcagg cggcgcaggg
      661 cctggttgtc gaccaccgcg ccacggctgg cgttgaccag ccaggtgccg ggacgcaggg
      721 cggccaggcg gggctcatcc agcaggtggc gggtcgggtg ttcgccgtcg cggttcagcg
      781 gagtgtgcag gctgatcacg tcggcctcgg ccagcagccg ttccagcgag acgaattcgc
      841 cgtccggctc gcgtgcctgg cgcggcggat cgcagaccag cactttccag cccagcccgc
      901 gcagcacctc caccagccgc ccgccgacct gcccggcgcc gaccacccca taggtgcgtt
      961 cggccaggtc ggcgccgcgc acctcggcca tcgccagcag gcagccgagc acgtagtcca
     1021 ccaccccgcg ggcgttgcag ccgggtgcgc tggaccaggc gatgccggcc tcggcgaagt
     1081 agtcgaggtc gaggtggtcg gtgccgatgg tgcaggtgcc gacgaagcgc accggcgagc
     1141 cggccagcgc cgcgcggctg acctcggtga ccgagcgcac cagcagcacg tcgacctcgg
     1201 ccagcgccgc gcggtcgatg gcgcgcccgg gcaagcggcg aatggagccc tggtcggcga
     1261 agaaggcgtc gaccacggga atgttttcat cggcgagaat acgcatatgt atatctcctt
     1321 cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca attcccctat
     1381 agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg acgcatcgtg
     1441 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
     1501 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     1561 gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg
     1621 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat
     1681 aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc gcggtatggc
     1741 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     1801 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     1861 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     1921 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     1981 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     2041 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     2101 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     2161 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     2221 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     2281 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     2341 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     2401 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     2461 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     2521 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     2581 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     2641 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     2701 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     2761 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     2821 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2881 taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga gccttcaacc
     2941 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     3001 tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg
     3061 accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc
     3121 acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg
     3181 ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg tcgttgagga
     3241 cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc
     3301 gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct
     3361 tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt
     3421 tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa
     3481 cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca
     3541 gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg
     3601 tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca
     3661 cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa
     3721 gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca
     3781 tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg
     3841 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3901 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     3961 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     4021 ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat accgcacaga
     4081 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     4141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4981 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5041 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5101 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5161 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5221 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5281 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5341 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     5401 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5461 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5521 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5581 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5641 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5701 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5761 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5821 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5881 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5941 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6001 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat
     6061 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     6121 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     6181 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     6241 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     6301 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     6361 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     6421 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6481 gcgccgctac agggcgcgtc ccattcgcca
//
