LOCUS       sequence-7407-ex        6180 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 6180)
  AUTHORS   Lee BI, Suh SW
  TITLE     Crystal structure of UDP-N-acetylglucosamine acyltransferase from 
            Helicobacter pylori.
  JOURNAL   Proteins. 2003 Nov 15. 53(3):772-4.
  PUBMED    14579368
REFERENCE   2  (bases 1 to 6180)
  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: "Proteins. 
            2003 Nov 15. 53(3):772-4."
COMMENT     Sequence Label: pET-21a(+)-LpxA
FEATURES             Location/Qualifiers
     source          1..6180
                     /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             984..1006
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1021..1045
                     /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(1045..1064)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1046..1064)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1148..1167)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1373..1450
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1451..2533
                     /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(1470..1489)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3342..3533
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3527..3549)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3635..3777
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3690..3709
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3792..3809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3963..4551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4043..4062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4722..5513,5514..5582))
                     /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     5345..5364
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5583..5687)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5714..6169)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5851..5872)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6063..6082
                     /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 gagttcctct tgtttttctt
      181 cgctagaata ttcgcttgac ttatacgcca cgcccctaga actctctaaa ataaaagagc
      241 aaatctcttt cacaaaaggg ttattggcat gctcttctaa ttctaattta gcgctctctc
      301 ttaaagaggg gatagggcgg aacaatcttt tataaagtgc ataaataaaa tcaatatcct
      361 tactctctaa taattggcgc atccggtggc ggtttaaccc cctaataaaa gcgcgattgc
      421 cctctacggt gcaataaggc ggcacgtctt tgcctaaagc gctcttaccg gctatcatgc
      481 accctttagc gatacgcaca aactgatgga tcgcggtaag accgccgata ttgacataat
      541 cgcctatttc aatatgacct gctaaagtta cgccattagc caaaatacaa tggcttccaa
      601 tcacgcaatc atgagcgaca tgcacataag ccatgagcag gtttttatcc ccaataaggg
      661 tttttttaat ccccccttca gtgccgggat ttatcatgca aaattcccga ataaggttgt
      721 cttctccaat aatcagttcg ctgtattcgc ccttatattt taaatcttga ggctgtgtgc
      781 ctagcacggc aaaagggaaa atttcggtgt ttttcccaac gaaagtatgc ccttgtaagg
      841 ttacattgtt gtggagtttc acgccttcat ctagtttgac gccatctcca atcacgcaaa
      901 attccccaat ctctacgccc ttattaatct ctgctttagg agagatgatg gctgtttttg
      961 caatcttact catcatatgt atatctcctt cttaaagtta aacaaaatta tttctagagg
     1021 ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag
     1081 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     1141 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     1201 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
     1261 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
     1321 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat
     1381 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     1441 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     1501 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     1561 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     1621 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     1681 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     1741 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     1801 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     1861 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     1921 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     1981 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     2041 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     2101 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     2161 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     2221 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     2281 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     2341 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     2401 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     2461 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     2521 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat
     2581 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     2641 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     2701 cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg
     2761 gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc
     2821 ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccccacggg
     2881 tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg
     2941 gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt
     3001 ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa
     3061 cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc
     3121 taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt
     3181 ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc
     3241 gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca
     3301 ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa
     3361 ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca
     3421 acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg
     3481 agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     3541 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     3601 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     3661 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     3721 ccatatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     3781 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3841 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3901 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3961 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4021 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4081 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4141 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4201 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4261 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4321 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4381 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     4441 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4501 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4561 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4621 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4681 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4741 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4801 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4861 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4921 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4981 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     5041 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5101 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5161 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5221 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5281 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5341 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5401 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5461 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5521 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5581 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5641 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5701 aaagtgccac ctgaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     5761 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5821 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5881 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5941 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     6001 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     6061 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     6121 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
