LOCUS       62056_2595        9446 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9446)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1402..2205
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        2346..2549
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2620..3315
                     /codon_start=1
                     /label=amilFP593
                     /note="amilFP593 is a basic (constitutively fluorescent)
                     red fluorescent protein published in 2007, derived from 
                     Acropora millepora."
                     /translation="MALSKHGLTKDMTMKYHMEGSVDGHKFVITGHGNGNPFEGKQTMN
                     LCVVEGGPLPFSEDILSATFDYGNRVFTEYPQGMVDFFKNSCPAGYTWHRSLLFEDGAV
                     CTTSADITVSVEENCFYHNSKFHGVNFPADGPVMKKMTTNWEPSCEKIIPVPRQGILKG
                     DIAMYLLLKDGGRYRCQFDTIYKAKSDPKEMPEWHFIQHKLTREDRSDAKNQKWQLVEH
                     AVASRSALPG"
     polyA_signal    3354..3578
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3625..3836
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3849..4117
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4173..4838
                     /codon_start=1
                     /label=CopGFP
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
                     /translation="LPAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             4908..4961
                     /codon_start=1
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4962..5558
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    5669..5801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    6054..6890
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(7425..7441)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7449..7465)
                     /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(7473..7503)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7518..7539)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7827..8415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8589..9446)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc caaggcctac tagtaacggc cgccagtgtg
     1381 ctggaattcg cccttggtac ctgctttctc tgaccagcat tctctcccct gggcctgtgc
     1441 cgctttctgt ctgcagcttg tggcctgggt cacctctacg gctggcccag atccttccct
     1501 gccgcctcct tcaggttccg tcttcctcca ctccctcttc cccttgctct ctgctgtgtt
     1561 gctgcccaag gatgctcttt ccggagcact tccttctcgg cgctgcacca cgtgatgtcc
     1621 tctgagcgga tcctccccgt gtctgggtcc tctccgggca tctctcctcc ctcacccaac
     1681 cccatgccgt cttcactcgc tgggttccct tttccttctc cttctggggc ctgtgccatc
     1741 tctcgtttct taggatggcc ttctccgacg gatgtctccc ttgcgtcccg cctccccttc
     1801 ttgtaggcct gcatcatcac cgtttttctg gacaacccca aagtaccccg tctccctggc
     1861 tttagccacc tctccatcct cttgctttct ttgcctggac accccgttct cctgtggatt
     1921 cgggtcacct ctcactcctt tcatttgggc agctccccta ccccccttac ctctctagtc
     1981 tgtgctagct cttccagccc cctgtcatgg catcttccag gggtccgaga gctcagctag
     2041 tcttcttcct ccaacccggg cccctatgtc cacttcagga cagcatgttt gctgcctcca
     2101 gggatcctgt gtccccgagc tgggaccacc ttatattccc agggccggtt aatgtggctc
     2161 tggttctggg tacttttatc tgtcccctcc accccacagt ggggcacgcg taagggcgaa
     2221 ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagtat tatgcccagt
     2281 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     2341 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     2401 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     2461 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     2521 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
     2581 gccatccacg ctgttttgac ctccatagaa gattctagaa tggcccttag taagcacggc
     2641 ctgactaagg acatgaccat gaagtaccat atggagggct cggtggacgg gcacaagttc
     2701 gtgattacgg gccacgggaa cggcaaccca ttcgagggga aacagacaat gaacctgtgt
     2761 gtggtggaag gcggacccct gcccttctcg gaggacatcc tgtccgcaac ctttgactac
     2821 ggcaatcgcg tgttcacaga gtatccacag ggtatggtgg atttctttaa gaacagctgt
     2881 cccgcaggat acacctggca ccgcagcctg ctcttcgagg acggggcggt gtgtaccacc
     2941 tccgctgata tcaccgtgtc cgtcgaggag aactgcttct accacaacag caaattccac
     3001 ggcgtgaact ttcctgccga cgggcctgtg atgaagaaaa tgactacgaa ttgggagccc
     3061 agctgcgaga aaatcatccc ggtccctcgt cagggaatct tgaagggcga catcgcaatg
     3121 tacctcctgc tcaaggacgg gggaagatac cgatgccagt tcgataccat ctacaaggca
     3181 aagtccgatc ccaaggagat gccagagtgg cacttcatcc agcacaaact cacccgcgag
     3241 gaccgctcag atgccaagaa ccagaagtgg caactggtcg agcacgccgt ggcatcccgt
     3301 agcgccctgc cagggtaatt taaacggcgc gccttcgaat ttaaatcgga tccctgtgcc
     3361 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3421 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3481 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3541 caatagcagg catgctgggg atgcggtggg ctctatggag atctgcggcc gcaaggatct
     3601 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     3661 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
     3721 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     3781 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga
     3841 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc
     3901 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg
     3961 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga
     4021 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg
     4081 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc
     4141 tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc gagtgccgca
     4201 tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag ggcaccccca
     4261 agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc ttcagcccct
     4321 acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac cccagcggct
     4381 acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc cgcatcgaga
     4441 agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag gccggccgcg
     4501 tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg atcttcaccg
     4561 acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc gataacgtgc
     4621 tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac agcttcgtgg
     4681 tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag aacgggggcc
     4741 ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg
     4801 agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct cagtcgtcca
     4861 attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag ggcagaggaa
     4921 gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag tacaagccca
     4981 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     5041 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     5101 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     5161 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     5221 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     5281 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     5341 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     5401 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     5461 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     5521 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct ctggattaca
     5581 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5641 acgctgcttt aatgcctttg tatcagttaa cttgtttatt gcagcttata atggttacaa
     5701 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5761 tggtttgtcc aaactcatca atgtatctta tcatgtctgg aattgactca aatgatgtca
     5821 attagtctat cagaagctca tctggtctcc cttccggggg acaagacatc cctgtttaat
     5881 atttaaacag cagtgttccc aaactgggtt cttatatccc ttgctctggt caaccaggtt
     5941 gcagggtttc ctgtcctcac aggaacgaag tccctaaaga aacagtggca gccaggttta
     6001 gccccggaat tgactggatt ccttttttag ggcccattgg tatggtgtac actactaggg
     6061 acaggattgg tgacagaaaa gccccatcct taggcctcct ccttcctagt ctcctgatat
     6121 tgggtctaac ccccacctcc tgttaggcag attccttatc tggtgacaca cccccatttc
     6181 ctggagccat ctctctcctt gccagaacct ctaaggtttg cttacgatgg agccagagag
     6241 gatcctggga gggagagctt ggcagggggt gggagggaag ggggggatgc gtgacctgcc
     6301 cggttctcag tggccaccct gcgctaccct ctcccagaac ctgagctgct ctgacgcggc
     6361 tgtctggtgc gtttcactga tcctggtgct gcagcttcct tacacttccc aagaggagaa
     6421 gcagtttgga aaaacaaaat cagaataagt tggtcctgag ttctaacttt ggctcttcac
     6481 ctttctagtc cccaatttat attgttcctc cgtgcgtcag ttttacctgt gagataaggc
     6541 cagtagccag ccccgtcctg gcagggctgt ggtgaggagg ggggtgtccg tgtggaaaac
     6601 tccctttgtg agaatggtgc gtcctaggtg ttcaccaggt cgtggccgcc tctactccct
     6661 ttctctttct ccatccttct ttccttaaag agtccccagt gctatctggg acatattcct
     6721 ccgcccagag cagggtcccg cttccctaag gccctgctct gggcttctgg gtttgagtcc
     6781 ttggcaagcc caggagaggc gctcaggctt ccctgtcccc cttcctcgtc caccatctca
     6841 tgcccctggc tctcctgccc cttccctaca ggtgttcctg gctctgctct ctcgagatgc
     6901 atgcgtcaat tttacgcaga ctatctttct agggttaatc tagctgcatc aggatcatat
     6961 cgtcgggtct tttttccggc tcagtcatcg cccaagctgg cgctatctgg gcatcgggga
     7021 ggaagaagcc cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt ttgccgagga
     7081 tgactgctgc tgcattgacg ttgagcgaaa acgcacgttt accatgatga ttcgggaagg
     7141 tgtggccatg cacgccttta acggtgaact gttcgttcag gccacctggg ataccagttc
     7201 gtcgcggctt ttccggacac agttccggat ggtcagcccg aagcgcatca gcaacccgaa
     7261 caataccggc gacagccgga actgccgtgc cggtgtgcag attaatgaca gcggtgcggc
     7321 gctgggatat tacgtcagcg aggacgggta tcctggctgg atgccgcaga aatggacatg
     7381 gataccccgt gagttacccg gcgggcgcgc ttggcgtaat catggtcata gctgtttcct
     7441 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7501 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7561 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7621 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7681 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7741 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7801 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7861 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7921 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7981 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     8041 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     8101 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8161 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8221 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     8281 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8341 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8401 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8461 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8521 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8581 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8641 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8701 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8761 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8821 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8881 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8941 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     9001 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     9061 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     9121 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     9181 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     9241 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     9301 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     9361 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9421 gcgacacgga aatgttgaat actcat
//
