LOCUS       sequence-348393-        5478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5478)
  AUTHORS   Yokouchi Y, Vogan KJ, Pearse RV, Tabin CJ
  TITLE     Antagonistic signaling by Caronte, a novel Cerberus-related gene, 
            establishes left-right asymmetric gene expression.
  JOURNAL   Cell. 1999 Sep 3. 98(5):573-83.
  PUBMED    10490097
REFERENCE   2  (bases 1 to 5478)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 1999 
            Sep 3. 98(5):573-83."
COMMENT     Sequence Label: pFLAG Caronte (CT#304)
FEATURES             Location/Qualifiers
     source          1..5478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1469..1492
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1502..2124
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1701..1720)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2148..2168)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2153..2482
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2333..2468
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2520..2539)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2521..2539)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2553..2569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2553..2569)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2566..2588)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2577..2593
                     /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)."
     promoter        complement(join(2601..2607,2608..2625,2626..2631))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2646..2667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2784..2801)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2955..3543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3035..3054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3714..4505,4506..4574))
                     /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"
     primer_bind     4337..4356
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4575..4679)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4706..5161
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4793..4812)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5003..5024
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5287..5309
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5301..5318
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5302..5318
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattcaag cttcgagcca tggcactgct cctccttcag ctgctagtgc tctcgtgtct
      661 tggagacaca gagccacagc cagattcaca gcaaaggaaa aggcggccac ttcagcacct
      721 tttctatttg gacagaaact tacttgaaag ccagagtttt catgagttgg tgggggaaaa
      781 cccagtaggt gttaaggaaa cccaggaaga gccaagcttt tttatagcat ttccacagac
      841 agcaggagag agtcagaagc aaggggagaa aaaaatgtcc agattcatcc ttcctaatgc
      901 agaactctat gcacaccaag atctgagaac ctgggcagca cccaaagaga tctctcctgt
      961 ggaaaacttc tctccatcct actattccaa caagagggat gttgaacctc cctatagaaa
     1021 agatgccaaa aaattctggg accatttcat gttaaggaaa aattcagctt ctgaagaggt
     1081 tgtcctgcca atcaagacca atgaaatgca ccaagaaact tgcagaaccc tgcctttttc
     1141 ccagagtgtt gctcatgaga gctgtgagaa ggtgatagtc cagaacaatc tatgttttgg
     1201 caagtgtagt tcctttcatg ttcctggtcc agatgatcgt ctttatacct tttgttctaa
     1261 atgcctgccc accaagtttt ccatgaagca cttggatctc aactgcacca gttctgtccc
     1321 agtggtcaaa aaagtcatga ttgtggaaga gtgcaactgt gagactcaga agatagaaga
     1381 tcctctgctt ggatctcgac agtcagattt tcttggaaat gtacctgagc acaatctgca
     1441 ggatatcagt cgacggatcc ggtaccaaga ttacaaggac gacgatgaca agtagctcga
     1501 ggggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     1561 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     1621 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     1681 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     1741 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     1801 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     1861 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     1921 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     1981 gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca
     2041 tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc
     2101 cactcagtag atgcctgttg aattgggtac gcggccagct tggctgtgga atgtgtgtca
     2161 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2221 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2281 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2341 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2401 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     2461 tggaggccta ggcttttgca aaaagctcct cgaggaactg aaaaaccaga aagttaattc
     2521 cctatagtga gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
     2581 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2641 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2701 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2761 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2821 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2881 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2941 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3001 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3061 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3121 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3181 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3241 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3301 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3361 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3421 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3481 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3541 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3601 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3661 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3721 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3781 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3841 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3901 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3961 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4021 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4081 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4141 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4201 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4261 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4321 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4381 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4441 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4501 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4561 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4621 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4681 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4741 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4801 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4861 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4921 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4981 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5041 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5101 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     5161 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     5221 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     5281 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctgatc tatacattga
     5341 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     5401 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     5461 caatatgacc gccatgtt
//
