LOCUS       40924_10111        6647 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6647)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6647)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             986..2014
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRTLDSETGAMVRLLE
                     DGD"
     promoter        complement(2199..2217)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2243..2467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2513..2941
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2955..3285
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3352..4143
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4320..4453
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4490..4506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4514..4530)
                     /label=lac operator
                     /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)."
     promoter        complement(4538..4568)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4583..4604)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4892..5480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5654..6511)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6512..6616)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggcacg
      901 agctggactt taagcaagca cctaaggaca tccaaaagcg taaacggatc cactgttaac
      961 ggccgcgggt gtgctggaat tcaccatgtc caatttactg accgtacacc aaaatttgcc
     1021 tgcattaccg gtcgatgcaa cgagtgatga ggttcgcaag aacctgatgg acatgttcag
     1081 ggatcgccag gcgttttctg agcatacctg gaaaatgctt ctgtccgttt gccggtcgtg
     1141 ggcggcatgg tgcaagttga ataaccggaa atggtttccc gcagaacctg aagatgttcg
     1201 cgattatctt ctatatcttc aggcgcgcgg tctggcagta aaaactatcc agcaacattt
     1261 gggccagcta aacatgcttc atcgtcggtc cgggctgcca cgaccaagtg acagcaatgc
     1321 tgtttcactg gttatgcggc ggatccgaaa agaaaacgtt gatgccggtg aacgtgcaaa
     1381 acaggctcta gcgttcgaac gcactgattt cgaccaggtt cgttcactca tggaaaatag
     1441 cgatcgctgc caggatatac gtaatctggc atttctgggg attgcttata acaccctgtt
     1501 acgtatagcc gaaattgcca ggatcagggt taaagatatc tcacgtactg acggtgggag
     1561 aatgttaatc catattggca gaacgaaaac gctggttagc accgcaggtg tagagaaggc
     1621 acttagcctg ggggtaacta aactggtcga gcgatggatt tccgtctctg gtgtagctga
     1681 tgatccgaat aactacctgt tttgccgggt cagaaaaaat ggtgttgccg cgccatctgc
     1741 caccagccag ctatcaactc gcgccctgga agggattttt gaagcaactc atcgattgat
     1801 ttacggcgct aaggatgact ctggtcagag atacctggcc tggtctggac acagtgcccg
     1861 tgtcggagcc gcgcgagata tggcccgcgc tggagtttca ataccggaga tcatgcaagc
     1921 tggtggctgg accaatgtaa atattgtcat gaactatatc cgtaccctgg atagtgaaac
     1981 aggggcaatg gtgcgcctgc tggaagatgg cgattagctc gagagtctca tcgccaatga
     2041 acgtcaggtg ttttggagaa ggtcacaccg cgcggagatg ttgaaacaaa gatctaactc
     2101 ttttgcaaca agatcaataa aataatgtta aaagcaaaaa aaaaaaaaaa aaaaaaaaaa
     2161 aaaaatctag atcgatagcg cgctagcccg ggccctattc tatagtgtca cctaaatgct
     2221 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2281 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2341 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2401 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2461 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2521 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2581 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2641 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     2701 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2761 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2821 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2881 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2941 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc
     3001 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3061 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3121 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3181 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3241 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3301 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3361 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3421 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3481 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3541 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3601 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3661 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3721 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3781 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3841 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3901 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3961 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4021 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4081 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4141 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4201 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4261 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4321 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4381 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4441 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4501 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4561 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4621 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4681 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4741 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4801 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4861 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4921 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4981 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5041 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     5101 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5161 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5221 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5281 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     5341 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5401 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5461 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5521 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5581 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5641 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5701 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5761 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5821 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5881 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5941 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6001 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6061 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6121 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6181 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6241 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6301 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6361 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6421 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6481 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6541 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6601 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
