LOCUS       Exported                6196 bp ds-DNA     circular SYN 08-MAY-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6196)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 25, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6196
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(367..1227)
                     /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(1228..1320)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1401..1989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2175..2317
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2419..2610)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3182..4264)
                     /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(4265..4342)
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4549..4566
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4576..4593
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4597..5316
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             5414..5446
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             5453..5470
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5636..5722
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5814..5841
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5909..5956
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      join(5993..6196,1..252)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
       61 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
      121 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
      181 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
      241 cgtttacaat ttctggcggc acgatggcat gagattatca aaaaggatct tcacctagat
      301 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
      361 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
      421 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
      481 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
      541 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
      601 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
      661 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
      721 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
      781 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
      841 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
      901 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
      961 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1021 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1081 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1141 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     1201 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatcatgat tgaagcattt
     1261 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     1321 taggtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1381 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1441 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1501 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     1561 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1621 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1681 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1741 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1801 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1861 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1921 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1981 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2041 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2101 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2161 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2221 cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     2281 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     2341 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2401 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     2461 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     2521 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     2581 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     2641 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     2701 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     2761 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     2821 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     2881 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     2941 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     3001 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     3061 gatcatgcta gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     3121 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     3181 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3241 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     3301 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     3361 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     3421 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca
     3481 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     3541 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     3601 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     3661 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     3721 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     3781 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     3841 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     3901 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     3961 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     4021 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     4081 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     4141 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     4201 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     4261 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     4321 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     4381 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     4441 tgcaaaattt atcaaaaaga gtgttgacta tactcagtgc cctgactatg atacttagat
     4501 tcatactcag tgccctgact atgaaggaga tataccatgg gcagcagcca tcatcatcat
     4561 catcacagca gcggcctggt gccgcgcggc agccatatgg tgagcaaggg cgaggagctg
     4621 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     4681 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     4741 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     4801 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     4861 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     4921 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     4981 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     5041 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     5101 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     5161 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     5221 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     5281 gggatcactc tcggcatgga cgagctgtac aagtaaggat ccgaattctg tacaggcgcg
     5341 cctgcaggac gtcgacggta ccatcgatac gcgttcgaag cttgcggccg cacagctgta
     5401 tacacgtgca agccagccag aactcgctcc tgaagaccca gaggatctcg agcaccacca
     5461 ccaccaccac taatgttaat taagttgggc gttcctaggc tgataaaaca gaatttgcct
     5521 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
     5581 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
     5641 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     5701 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
     5761 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
     5821 catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac
     5881 attcaaatat gtatccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     5941 cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc
     6001 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6061 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6121 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6181 cggcacctcg acccca
//
