LOCUS       Exported                5946 bp ds-DNA     circular SYN 13-AUG-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5946)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 28, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5946
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        15..81
                     /gene="cspA"
                     /label=cspA promoter
                     /note="promoter of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     protein_bind    84..100
                     /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)."
     5'UTR           122..255
                     /gene="cspA"
                     /label=cspA 5'UTR
                     /note="5'UTR of the E. coli cold shock protein cspA gene 
                     (Mitta et al., 1997)"
     primer_bind     197..218
                     /label=090 pColdI seq-F
     primer_bind     197..218
                     /label=108 T7_SDseq_EGFP-F
     primer_bind     complement(226..255)
                     /label=pCold-R
     primer_bind     241..280
                     /label=HisSBP4A1-F
     CDS             271..285
                     /codon_start=1
                     /label=His6
                     /translation="HHHHH"
     CDS             286..408
                     /codon_start=1
                     /label=SBP
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREPSGGG"
     misc_feature    286..408
                     /label=SBP
                     /note="""/ApEinfo_fwdcolor=#00ff00"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=green"
     primer_bind     286..302
                     /label=043 FRTTO kozac SBP-F
                     /note="pcDNA5 FRT/TO
                       BamHcut＋kozac+SBP"
     5'UTR           286
                     /label=Kozak
                     /note=" [Split]"
     misc_feature    409..1836
                     /label=DDX3X core
     primer_bind     1840..1864
                     /label=pCold-F
     primer_bind     1876..1891
                     /label=19.2.1 PcoldI-HS backbone F primer
     3'UTR           1907..2051
                     /gene="cspA"
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene 
                     (Mitta et al., 1997)"
     primer_bind     complement(1923..1943)
                     /label=091 pColdI seq-R
     rep_origin      complement(2250..2705)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2805..2909
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2910..3770
                     /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"
     rep_origin      3941..4529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4701..5783)
                     /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(5784..5861)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa
       61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat
      121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt
      181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt
      241 aagaggtaat acaccatggg cagcagccat catcatcatc atcacgacga gaaaaccacc
      301 ggctggcggg gaggccacgt ggtggaaggg ctggcaggcg agctggaaca gctgcgggcc
      361 agactggaac accaccccca gggccagaga gagcctagcg gcggaggaga tgaagatgat
      421 tggtcaaaac cactcccacc aagtgaacgc ttggaacagg aactcttttc tggaggcaac
      481 actgggatta attttgagaa atacgatgac attccagttg aggcaacagg caacaactgt
      541 cctccacata ttgaaagttt cagtgatgtt gagatgggag aaattatcat gggaaacatt
      601 gagcttactc gttatactcg cccaactcca gtgcaaaagc atgctattcc tattatcaaa
      661 gagaaaagag acttgatggc ttgtgcccaa acagggtctg gaaaaactgc agcatttctg
      721 ttgcccatct tgagtcagat ttattcagat ggtccaggcg aggctttgag ggccatgaag
      781 gaaaatggaa ggtatgggcg ccgcaaacaa tacccaatct ccttggtatt agcaccaacg
      841 agagagttgg cagtacagat ctacgaggaa gccagaaaat tttcataccg atctagagtt
      901 cgtccttgcg tggtttatgg tggtgccgat attggtcagc agattcgaga cttggaacgt
      961 ggatgccatt tgttagtagc cactccagga cgtctagtgg atatgatgga aagaggaaag
     1021 attggattag acttttgcaa atacttggtg ttagatgaag ctgatcggat gttggatatg
     1081 gggtttgagc ctcagattcg tagaatagtc gaacaagata ctatgcctcc aaagggtgtc
     1141 cgccacacta tgatgtttag tgctactttt cctaaggaaa tacagatgct ggctcgtgat
     1201 ttcttagatg aatatatctt cttggctgta ggaagagttg gctctacctc tgaaaacatc
     1261 acacagaaag tagtttgggt ggaagaatca gacaaacggt catttctgct tgacctccta
     1321 aatgcaacag gcaaggattc actgacctta gtgtttgtgg agaccaaaaa gggtgcagat
     1381 tctctggagg atttcttata ccatgaagga tacgcatgta ccagcatcca tggagaccgt
     1441 tctcagaggg atagagaaga ggcccttcac cagttccgct caggaaaaag cccaatttta
     1501 gtggctacag cagtagcagc aagaggactg gacatttcaa atgtgaaaca tgttatcaat
     1561 tttgacttgc caagtgatat tgaagaatat gtacatcgta ttggtcgtac gggacgtgta
     1621 ggaaaccttg gcctggcaac ctcattcttt aacgagagga acataaatat tactaaggat
     1681 ttgttggatc ttcttgttga agctaaacaa gaagtgccgt cttggttaga aaacatggct
     1741 tatgaacacc actacaaggg tagcagtcgt ggacgttcta agagtagcag atttagtgga
     1801 gggtttggtg ccagagacta ccgacaaagt agcggttgac atatggagct cggtaccctc
     1861 gagggatccg aattcaagct tgtcgacctg cagtctagat aggtaatctc tgcttaaaag
     1921 cacagaatct aagatccctg ccatttggcg gggatttttt tatttgtttt caggaaataa
     1981 ataatcgatc gcgtaataaa atctattatt atttttgtga agaataaatt tgggtgcaat
     2041 gagaatgcgc aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2101 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2161 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2221 agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt
     2281 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2341 cccttataaa tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa
     2401 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2461 cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa
     2521 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2581 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2641 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2701 cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     2761 aaataccgca tcaggcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2821 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2881 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2941 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3001 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3061 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3121 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3181 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3241 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3301 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3361 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3421 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3481 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3541 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3601 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3661 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3721 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3781 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3841 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3901 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3961 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4021 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4081 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4141 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4201 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4261 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4321 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4381 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4441 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4501 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4561 tggccttttg ctggcctttt gctcacatag tcatgccccg cgcccaccgg aaggagctga
     4621 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     4681 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4741 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     4801 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     4861 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     4921 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     4981 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     5041 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     5101 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     5161 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     5221 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     5281 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     5341 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     5401 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     5461 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     5521 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     5581 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     5641 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     5701 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     5761 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     5821 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     5881 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     5941 cgccgc
//
