LOCUS       sequence-7779-ex        5898 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 5898)
  AUTHORS   Kang JY, Lee HH, Yoon HJ, Kim HS, Suh SW
  TITLE     Overexpression, crystallization and preliminary X-ray 
            crystallographic analysis of phosphopantetheine adenylyltransferase 
            from Enterococcus faecalis.
  JOURNAL   Acta Crystallograph Sect F Struct Biol Cryst Commun. 2006 Nov 1. 
            62(Pt 11):1131-3.
  PUBMED    17077496
REFERENCE   2  (bases 1 to 5898)
  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 Nov 1. 62(Pt 
            11):1131-3."
COMMENT     Sequence Label: EF2451_pET21a(+)
FEATURES             Location/Qualifiers
     source          1..5898
                     /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"
     CDS             complement(662..694)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     RBS             702..724
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    739..763
                     /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(763..782)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(764..782)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(866..885)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1091..1168
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1169..2251
                     /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(1188..1207)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3060..3251
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3245..3267)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3353..3495
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3408..3427
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3510..3527)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3681..4269)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3761..3780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4440..5231,5232..5300))
                     /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     5063..5082
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5301..5405)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5432..5887)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5569..5590)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5781..5800
                     /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 gaggctccag tcattcttct
      181 tttgttttaa cgcatgataa atattgggag gaagatagtc agacacatcg cccccaaaac
      241 gtaaaacttc ttttagtaaa ctggaactta catgtgcata tggttcttct gctaatagaa
      301 aaaccgtttc aatttctggc gccagatgtt gattcatttt agcaatgtct ttctcgtact
      361 cgtagtcttt gacgtttcta atgccacgaa tcaaaaaatt agcccctaat gattttgcac
      421 tttccacagt taactgggtt tcttgcataa taacacgcac atttggcatt tctttagttg
      481 cttcttcaat taaatatttt ttctcttctg gcgtaaataa tgtttgctta cttgtattta
      541 taaaaacgcc aataatgact tcgtcaaata acttcgcact gcgttcaatc aaatttaaat
      601 gaccatttgt catcggatca aaacttccgg gaaatagagc aattttacgc atggatccgc
      661 gacccatttg ctgtccacca gtcatgctag ccatatgtat atctccttct taaagttaaa
      721 caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat
      781 taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc
      841 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
      901 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
      961 gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc
     1021 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
     1081 cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
     1141 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     1201 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     1261 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     1321 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     1381 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     1441 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     1501 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     1561 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     1621 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     1681 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     1741 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     1801 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     1861 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     1921 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     1981 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     2041 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     2101 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     2161 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     2221 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
     2281 ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     2341 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     2401 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
     2461 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
     2521 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
     2581 gcatggcggc cccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct
     2641 ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg
     2701 actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg
     2761 tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca
     2821 tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc
     2881 attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct
     2941 cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct
     3001 ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag
     3061 tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa
     3121 cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc
     3181 ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg
     3241 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     3301 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     3361 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     3421 gattgtactg agagtgcacc atatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     3481 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3541 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3601 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3661 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3721 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3781 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3841 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3901 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3961 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4021 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4081 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     4141 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4201 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4261 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4321 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4381 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4441 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4501 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4561 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4621 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4681 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4741 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4801 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4861 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4921 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4981 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5041 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5101 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5161 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5221 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5281 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5341 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5401 ccgcgcacat ttccccgaaa agtgccacct gaaattgtaa acgttaatat tttgttaaaa
     5461 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5521 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5581 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5641 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5701 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5761 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5821 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     5881 ggcgcgtccc attcgcca
//
