LOCUS       Exported                7307 bp DNA     circular SYN 10-SEP-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7307)
  AUTHORS   11111111
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        187..205
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    206..230
                     /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)."
     CDS             287..304
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             311..1795
                     /codon_start=1
                     /gene="nusA"
                     /product="transcription elongation protein (N utilization 
                     substance protein A)"
                     /label=NusA
                     /note="highly soluble in E. coli"
                     /translation="MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
                     RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
                     ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
                     REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
                     IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
                     QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
                     VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
                     EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
                     LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     CDS             1811..1828
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1838..1882
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1898..1915
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1934..1948
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             2060..2092
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             2099..2116
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2282..2368
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2460..2487
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      2555..2602
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2639..3094
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(3209..4069)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4070..4162)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4243..4831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5017..5156
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(5258..5449)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6021..7103)
                     /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"
     promoter        complement(7104..7181)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
       61 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
      121 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg
      181 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
      241 taattttgtt taactttaag aaggagatat acatatgggc agcagccatc accaccatca
      301 ccattctagt atgaacaaag aaattttggc tgtagttgaa gccgtatcca atgaaaaggc
      361 gctacctcgc gagaagattt tcgaagcatt ggaaagcgcg ctggcgacag caacaaagaa
      421 aaaatatgaa caagagatcg acgtccgcgt acagatcgat cgcaaaagcg gtgattttga
      481 cactttccgt cgctggttag ttgttgatga agtcacccag ccgaccaagg aaatcaccct
      541 tgaagccgca cgttatgaag atgaaagcct gaacctgggc gattacgttg aagatcagat
      601 tgagtctgtt acctttgacc gtatcactac ccagacggca aaacaggtta tcgtgcagaa
      661 agtgcgtgaa gccgaacgtg cgatggtggt tgatcagttc cgtgaacacg aaggtgaaat
      721 catcaccggc gtggtgaaaa aagtaaaccg cgacaacatc tctctggatc tgggcaacaa
      781 cgctgaagcc gtgatcctgc gcgaagatat gctgccgcgt gaaaacttcc gccctggcga
      841 ccgcgttcgt ggcgtgctct attccgttcg cccggaagcg cgtggcgcgc aactgttcgt
      901 cactcgttcc aagccggaaa tgctgatcga actgttccgt attgaagtgc cagaaatcgg
      961 cgaagaagtg attgaaatta aagcagcggc tcgcgatccg ggttctcgtg cgaaaatcgc
     1021 ggtgaaaacc aacgataaac gtatcgatcc ggtaggtgct tgcgtaggta tgcgtggcgc
     1081 gcgtgttcag gcggtgtcta ctgaactggg tggcgagcgt atcgatatcg tcctgtggga
     1141 tgataacccg gcgcagttcg tgattaacgc aatggcaccg gcagacgttg cttctatcgt
     1201 ggtggatgaa gataaacaca ccatggacat cgccgttgaa gccggtaatc tggcgcaggc
     1261 gattggccgt aacggtcaga acgtgcgtct ggcttcgcaa ctgagcggtt gggaactcaa
     1321 cgtgatgacc gttgacgacc tgcaagctaa gcatcaggcg gaagcgcacg cagcgatcga
     1381 caccttcacc aaatatctcg acatcgacga agacttcgcg actgttctgg tagaagaagg
     1441 cttctcgacg ctggaagaat tggcctatgt gccgatgaaa gagctgttgg aaatcgaagg
     1501 ccttgatgag ccgaccgttg aagcactgcg cgagcgtgct aaaaatgcac tggccaccat
     1561 tgcacaggcc caggaagaaa gcctcggtga taacaaaccg gctgacgatc tgctgaacct
     1621 tgaaggggta gatcgtgatt tggcattcaa actggccgcc cgtggcgttt gtacgctgga
     1681 agatctcgcc gaacagggca ttgatgatct ggctgatatc gaagggttga ccgacgaaaa
     1741 agccggagca ctgattatgg ctgcccgtaa tatttgctgg ttcggtgacg aagcgactag
     1801 tggttctggt catcaccatc accatcactc cgcgggtaaa gaaaccgctg ctgcgaaatt
     1861 tgaacgccag cacatggact cgccaccgcc aactggtctg gtcccccggg gcagcgcggg
     1921 ttctggtacg attgatgacg acgacaagag tccggagctc gtggatccga attctgtaca
     1981 ggcgcgcctg caggacgtcg acggtaccat cgatacgcgt tcgaagcttg cggccgcaca
     2041 gctgtataca cgtgcaagcc agccagaact cgctcctgaa gacccagagg atctcgagca
     2101 ccaccaccac caccactaat gttaattaag ttgggcgttc ctaggctgat aaaacagaat
     2161 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     2221 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     2281 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     2341 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     2401 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     2461 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     2521 aaatacattc aaatatgtat ccgctgagca ataactagca taaccccttg gggcctctaa
     2581 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     2641 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2701 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2761 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2821 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2881 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2941 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3001 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3061 gcgaatttta acaaaatatt aacgtttaca atttctggcg gcacgatggc atgagattat
     3121 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3181 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3241 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3301 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3361 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3421 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3481 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3541 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3601 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3661 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3721 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3781 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3841 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3901 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3961 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4021 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4081 ttcaatcatg attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4141 tgtatttaga aaaataaaca aataggtcat gaccaaaatc ccttaacgtg agttttcgtt
     4201 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4261 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4321 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4381 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4441 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4501 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4561 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4621 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4681 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4741 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4801 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4861 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4921 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4981 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     5041 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
     5101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     5161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     5341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     5401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     5461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     5521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     5581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     5641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     5701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     5761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     5821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     5881 tcctcaacga caggagcacg atcatgctag tcatgccccg cgcccaccgg aaggagctga
     5941 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     6001 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6061 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     6121 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     6181 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     6241 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     6301 atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     6361 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     6421 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     6481 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     6541 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     6601 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     6661 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     6721 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     6781 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     6841 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     6901 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     6961 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     7021 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     7081 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     7141 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     7201 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     7261 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccc
//
